Raffaella Leoci
Animal by-products (ABPs):
origins, uses, and European regulations
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Forewords
About half of each animal bred for meat, eggs and milk production, is not used
for human consumption. A part of that unused half is subjected to rendering
processes, resulting in many useful products. In the past industrial applications
have used leather and skins, wools, fat and fatty acids as a feedstock, whereas
relatively few applications were developed for animal by-products beyond adhesives and
fertilizers. During the last decades the demand for recovering by-products for these ap-
plications has declined with the increase in use of widely available petroleum/synthetic-
based products. A greater focus on renewable raw materials, however, has currently been
reported, putting the focus on co-products otherwise destined for disposal.
In order to add value to animal hide and internal organs by-products, the meat industry is
using science and research to add a value far beyond the usual profitability. It is necessary to
employ up-to-date research tools to study these co-products for their properties, in order to
develop new technological applications, and to search for medical, cosmetic or other indus-
trial applications.
Leoci, in the present study, equipped with a large number of references (over 500), reviews
the current uses of the various components of animal by-products— such as bile, blood, bones,
brains, fats and fatty acids, glands (adrenal, kidney, liver, pancreas, etc.), hides, hearts, intes-
tines, lungs, ovaries, stomachs, trachea, etc.— to show the "state of the art" of this science so
that interested readers (operators of slaughter, transport and disposal of the unusable parts)
and scholars may know in which direction we need to work and study in order to undertake
further research for recovering additional useful components. The animal by-products are to
be considered a mine of substances and compounds, still in large part undiscovered.
In the meantime, it should be noted, again warns Leoci, that the unusable parts wasted
need to be handled with caution to avoid problems of environmental pollution and health
hazards. Of course, the field of waste management is governed by numerous laws and regu-
lations that must be observed to avoid problems and penalties. Leoci examines and lists the
rules governing the management sector of the animal by-products, both that of waste (from
the collection and transport to the recovery and/or disposal) and the overlapping and inter-
secting rules of animal by-product waste management in Europe and in Italy, highlighting
the rules that cause problems.
Prof. Gaetano Vitale Celano
Department of Veterinary Medicine
Section of Food Inspection
University of Bari "Aldo Moro", Bari, Italy
Ban, October, 2013
In this remarkable work Leoci, stepping outside her usual field of animal reproduction
research, provides an overview of the state of the art of Animal By-Product (ABP)
utilization. Animal slaughter currently generates a large amount of waste. Despite the
progress over years past and recommendations of recent studies, approximately 50%
of the weight of a slaughtered animal finds no use at all and is disposed of in landfills or, at
most, used as a component in fertilizer. It's an unacceptable practice and a waste of renew-
able resources.
Among the various ABPs, greater attention needs to be paid to the rare recoveries and uses of
animal's reproductive organs, which, in this paper, Leoci gives notice. These organs are rich
in enzymes, hormones, vitamins, estrogens and countless other beneficial compounds that
can address deficiencies and dysfunctions in both the animal and human kingdoms. How-
ever, further studies are required to hone accuracy and to find and define the appropriate
application for the countless substances present in the animal reproductive organs. Studies
and research is necessary not only to identify additional probable use in the pharmaceutical
industry, but also and above all to better understand the complex mechanisms of cell repro-
duction, which until now is not completely clear. It is important to note that initial results
in this area of cell reproduction were obtained by the study of sea urchin eggs.
This book will therefore be useful not only to scholars, providing a wide scope of biblio-
graphical data (or notes) about relevant and recent research, but also to the workers and
management of the slaughtering industry, possibly encouraging the industry to fund further
studies in the field and thereby create an opportunity to reap economic returns thanks to the
increased use of ABPs. It will also be useful to those who are concerned with European rules
and regulations in the slaughtering industry and whose understanding of the issues will be
deepened by the information.
Prof. Giovanni Michele Lacalandra
Department of Emergency and Organ Transplantation (DETO)
Section of Veterinary Clinic and Animal Production
University of Bari "Aldo Moro", Bari, Italy
Bari, October 2013
Preface
Animal by-products (ABPs) arise mainly during the slaughter of animals during
the production of animal origin products (such as dairy, fibre, leather products),
as the result of disease control measures, and in the course of the disposal of dead
animals. Past and recent crises related to outbreaks of foot-and-mouth disease, the
spread of transmissible spongiform encephalopathies (TSE) and the occurrence of hazard-
ous chemicals (e.g. compounds of lead, mercury, chromium, PCBs, dioxins, etc.) in feeding
stuffs have shown the potential risk to public and animal health and the environment. This
risk needs to be adequately controlled, either by directing such products towards safe means
of disposal or by using them for different purposes, provided that strict conditions are ap-
plied which minimize the health risks involved. On the other hand the disposal of all ABPs
is not a realistic option, as it would lead to unsustainable costs for producers and consum-
ers and risks for the environment. Conversely, there is a clear interest for all citizens that,
provided the health risks are minimized, a wide range of ABPs can be safely used for various
applications in a sustainable manner.
A lot of ABPs are commonly used in important productive sectors, such as in the phar-
maceutical, feed, wool and leather industries but, notwithstanding, new technologies have
widened the possible use of ABPs and derived products. Consequently a wide range of ABPs
are not utilized and are destined to disposal.
This research first exposes the industry's "state of the art", namely what parts of ABPs are
used to produce more food and useful products for various industries besides the traditional
uses of fats and acids to produce energy or soaps; of hides and skins to produce footwear,
articles of apparel, gelatines, and glues; and of bones to produce fertilizers, special ceramics,
and glues. In addition, it shows there are many industries that use bile, blood, and glands to
produce a long list of pharmaceuticals. It also shows that many other by-products compo-
nents have yet to be studied in order to find suitable uses.
However, by eliminating all components used in various industries, including the food in-
dustry, there still remains a significant portion of ABPs that must be disposed. Part II men-
tions various possible disposal systems now in use: from sanitary landfills, to incineration
with or without energy recovery, to anaerobic digestion in order to produce biogas.
In the European Union the whole sector is governed by rules and laws that can be divided
into two branches: those governing the production, collection, transport and use of ABPs
that are usable and those governing the collection, transport and disposal of ABPs that can
not be used, since they are considered waste. In addition, Part II examines the various Euro-
pean rules (Regulations, Directives, etc.), adopted by all member nations and those specific
to enforcement in Italy. It has been noted that some of the same ruling applications are not
common to the two governing spheres. Some judgments of the Italian Constitutional Court,
make it possible to avoid a few mistakes and related penalties required by the EU, but it
would be appropriate to call on legislators to eliminate that uncertainty.
Raffaella Leoci
Department of Emergency and Organ Transplantation (DETO)
Section of Veterinary Clinic and Animal Production
University of Bari "Aldo Moro"
Bari, September 2013
Forewords
Preface
CONTENTS
3
9
PART I - Current uses of ABPs
1. Introduction 15
2. Animal by-products (ABPs): what are they? 17
3. Historical uses of ABPs 19
4 . Current uses of AB Ps 21
4.1 Bile 25
4.2 Blood 26
Edible blood (Food) 27
£j (blood meal) 43
Fertilizers 44
d) Scientific use or research 45
e) Laboratory uses 45
f) Pharmaceutical uses 50
Veterinary biological uses 55
/rj Industrial uses 56
4.3 Bones 61
4.4 Brains and spinal cords 62
4.5 Fats and fatty acids 64
4.6 Glands 68
a) Adrenal 68
b) Kidney 69
c) Liver 71
d) Pancreas 72
e) Pituitary glands 75
/) Spleen 77
g) Thymus 78
Thyroid 80
4.7 Hides and skins 83
4.8 Hairs and wools, nails, horns, feathers, hooves 86
4.9 Hearts 90
4.10 Intestines 92
4.11 Lungs 99
4.12 Meat and bone meals 99
4.13 Ovaries 101
4.14 Proteins 102
a) Protein hydrolysis products 102
b) Proteins for plastics 102
c) Proteins as adhesives 103
d) Protein surfactants 104
4.15 Stomachs and tripes 104
a) Ruminant stomach as food 105
b) Rumen and rennet 106
4.16 Trachea 108
a) Chondroitin sulphate 108
PART II - Treatment and legislation on ABPs and waste
5. Collection and treatment plants for ABPs 115
5.1 Rendering plants: handling and storage plants 115
5.2 Incinerators/Co-incineration Plants 117
5.3 Landfill 119
5.4 Anaerobic digestion (biogas) plants 119
5.5 Composting 122
5.6 Pet food plants 123
6. EU rules on animal by-products (ABPs) 125
6.1 Regulation (EC) 1069/2009 and Regulation (EU) 142/2011 134
7. EU rules on waste from ABPs 145
7.1 Waste Directive 145
7.2 Landfill Directive 148
7.3 Packaging and Packaging Waste Directive 150
7.4 End of Life Vehicles Directive 150
7.5 Waste Incineration Directive 151
7.6 Other Directives 151
8. Italian rules on ABPs and on waste from ABPs 151
8.1 Conclusions 158
References 161
EU and Italian Rules 189
PARTI
Current uses ofABPs
Animal by-products (ABPs): origins, uses, and European regulations
1. Introduction
Until 1986 the use of meals of animal origin as a protein supplement in cattle had no
restriction. In that year, the first cases 1 of Bovine Spongiform Encephalopathy (BSE) were
identified in GB. Many studies suggested that the first cases occurred around April 1985. 2
BSE, commonly known as mad cow disease, is a fatal neurodegenerative disease (encepha-
lopathy) in cattle 3 that causes a spongy degeneration in the brain and spinal cord. The disease
has become known to the public as Mad Cow Disease (MCD). The BSE belongs to a group
of diseases called "transmissible spongiform encephalopathies" (TSE), affecting different ani-
mal species, including humans. 4
The occurrence of the disease, caused by a strange pathogenic protein known as "prion"
or more commonly as an "unconventional infectious agent", was attributed to the use of ani-
mal meal as protein supplement in cattle, especially in the United Kingdom, where the rules,
on the high temperature treatment of animal by-products, were much less restrictive than
in other countries. The use of animal by-products (ABP) 5 of sick cattle, in the production of
meat meal and bone meal for animal feed, brought on the onset of BSE in cattle. Maternal
transmission was recently considered possible. 6
The existence of prions was discovered by Stanley Prusiner, 7 1997 Nobel Prize in Medi-
cine, who added these agents to the list of well-known infectious substances including
1. Wells G. A. H., A. C. Scott, C. T. Johnson, R. F. Gunning, R. D. Hancock, M. Jeffrey, M. Dawson,
R. Bradley (1987), "A novel progressive spongiform encephalopathy in cattle", Veterinary Record, vol.
121(18): 419-420.
2. Wilesmith J. V. (1991), "Epidemiology of bovine spongiform encephalopathy", Seminars in Virol-
ogy, vol. 2: 239-245.
3. In the UK they are not immediately understood the link between the feeding of cattle with meat
and bone meal and the rise of BSE. Refer for more news: Packer R. (2006), The Politics of BSE, Pal-
grave Macmillan, London; Wilesmith J. W, G. A. H. Wells, J. B. M. Ryan, D. Gavier-Widen, M. M.
Simmons (1997), "A cohort study to examine maternally-associated risk factors for bovine spongiform
encephalopathy", Veterinary Record, vol. 141:239-243.
4. See among others: Scoccia E., M. De Curtis, M. Biagetti, L. Faccenda, C. Maresca (2010), "Ence-
falopatia spongiforme bovina: excursus epidemiologico 2003-2008 - Bovine spongiform encephalopa-
thy: epidemiological overview 2003-2008", Sanita Pubblica Veterinaria, vol. 63 (12). ( http://indice.
spvet.it#500) .
5. "Animal by-Products (ABPs)" can be defined as animal carcasses, parts of carcasses or products of
animal origin (including ova, embryos and semen, sometimes glands, bloods, etc.) that are not in-
tended for human consumption.
6. Wilesmith J. W, G. A. H. Wells, J. B. M. Ryan, D. Gavier-Widen, M. M. Simmons (1997), Veteri-
nary Record, ibid.: 239-243.
7. S. B. Prusiner was Professor of Neurology at the University of California, San Francisco where he has
worked since 1972. From 1969-72, he served in the US Public Health Service at the National Institutes
of Health. He is the recipient of numerous prizes, including the Nobel Prize in Physiology or Medicine
(1997). Prusiner's work helped the world to understand more about Alzheimer's and Mad Cow disease
through his discovery of the prion, a disease-causing agent like bacteria or viruses (See: Prusiner S.
B. (1991), "Molecular Biology of Prion Desease", Science - New Series 1991, vol. 252 (5012): 1515-
1522). Prions cause transmissible and genetic neurodegenerative diseases, including scrapie and bovine
spongiform encephalopathy of animals and Creutzfeldt-Jakob and Gerstmann-Straussler-Scheinker
diseases of humans. The prion protein can manifest itself as two proteins, one an innocent "Dr. Jekyll"
character, while the other, dangerous "Mr Hyde" protein causes disease and death.
15
Raffaella Leoci
bacteria, viruses, fungi and parasites. Prions exist normally as innocuous cellular proteins;
however, they possess an innate capacity to convert their structures into highly stabile con-
formations that ultimately result in the formation of harmful particles, the causative agents
of several deadly brain diseases of the dementia type in humans and animals. Prion diseases
may be inherited, laterally transmitted, or occur spontaneously.
While a neurology resident, Prusiner was in charge of a patient who died of a rare fatal
degenerative disorder of the brain called "Creutzfeldt-Jakob Disease" 8 (vCJD o nvCJD).
Prusiner became intrigued by this little-known class of neurodegenerative disorders, the
spongiform encephalopathies that caused progressive dementia and death in humans and
animals. In 1974 he set up a laboratory to study scrapie, a related disorder of sheep, and in
1982 he claimed to have isolated the scrap ie-causing agent. He claimed that this pathogenic
agent, which he named "prion," was unlike any other known pathogen, such as a virus or
bacterium, because it consisted only of protein and lacked the genetic material contained
within all life-forms that is necessary for replication.
When first published, the prion theory met with much criticism, but it became widely
accepted by the mid-1990s. In 1996, when a new variant of "Creutzfeldt-Jakob Disease"
emerged in Great Britain, Prusiner's research was the focus of national attention. Fears
abounded that the new variant of the disease might be linked to "mad cow" disease, a brain
disorder that first appeared in British cattle a decade earlier. Some evidence suggested that
the mad cow prion might have jumped species, infecting humans who consumed beef con-
taminated with the infectious agent. Because mad cow disease was believed to have been
caused when the agent that causes scrapie in sheep was transmitted to cattle in feed, there
was precedent for species-jumping events to occur. Prusiner's research also could have sig-
nificant implications for such disorders as Alzheimer disease and Parkinson disease, which
seemed to share certain characteristics with the diseases caused by prions.
The leading scientific theory at this time maintained that CJD and the other TSEs were
caused by a type of prion. 9
8. Creutzfeldt-Jakob disease (CJD) is a rare, degenerative, invariably fatal brain disorder. It affects
about one person in every one million people per year worldwide. CJD belongs to a family of human
and animal diseases known as the transmissible spongiform encephalopathies. Spongiform refers to the
characteristic appearance of infected brains, which become filled with holes. The causes and origins
of Creutzfeldt-Jakob disease are not yet known. Some researchers believe an unusual "slow virus" or
another organism causes CJD. However, they have never been able to isolate a virus or other organism
in people with the disease. Furthermore, the agent that causes CJD has several characteristics that are
unusual for known organisms such as viruses and bacteria. It is difficult to kill: "it survives" or better,
it stays active after 30 minutes of immersion in boiling water, it is resistant to UV, to concentrated
formaldehyde, to phenol. "Survives" because maybe it is not a living organism, but a protein "contain-
ing" DNA. It does not appear to contain any genetic information in the form of nucleic acids (DNA
or RNA), and it usually has a long incubation period before symptoms appear. In some cases, the
incubation period may be as long as 50 years.
9. Intriguingly, the infectious prion protein is made by the host, and its amino acid sequence is identi-
cal to a normal host protein. Moreover, the prion and normal forms of the protein are indistinguish-
able in their posttranslational modifications. The only difference between them appears to be in their
folded three-dimensional structure. The misfolded prion protein tends to aggregate, and it has the re-
markable capacity to cause the normal protein to adopt its misfolded prion conformation and thereby
to become infectious. This ability of the prion to convert the normal host protein to misfolded prion
protein is equivalent to the prion's having replicated itself in the host. If eaten by another susceptible
16
Animal by-products (ABPs): origins, uses, and European regulations
Kuru is a human prion disease, very similar to BSE that was spread from one person
to another by ritual mortuary practices in New Guinea. However, the two Nobel Laureates,
Gajdusek 10 and Prusiner, never found an explanation of the unusual features of prion. Ad-
ditionally, biologist and Nobel Laureate James Watson" called prion the strangest thing in
molecular biology. To date, its chemical structure remains unknown and no one is certain of
having ever seen prion under a microscope.
2. Animal by-products (ABPs): what are they?
Slaughtered bred animals are commonly divided in two typical fractions: animal prod-
ucts used for food and not destined for food (edible and inedible ABPs). The amount of
by-products not destined for food is what slaughterhouses refer to as their "5th quarter".
Proportion of meat and by-products
(estimates)
Slaughtered animal
For human consumption (~%)
ABPs (~%)
(edible and inedible)
Cattle
55
45
Broilers
70
30
Pig
60
40
Sheep/goat
50
50
The above proportions are not valid for all countries. For instance, in India cattle are
100% inedible, while in Muslim countries pigs are 100% inedible. Even the proportions
may vary according to many factors (breeds, breeding conditions, feeding, etc.). Regarding
the ABPs, some parts (for instance livers, blood, lungs, etc.) are edible in some countries and
inedible in other countries, depending on eating habits. Anyway, inedible by-products in-
clude all slaughter by-products which are obviously not fit for human consumption: leathers,
feathers, bristles and horns or materials that have been declared not fit for human consump-
tion after a veterinary inspection (infected meats, injured animals, meat with inflammation
or bad smells, etc.). Inedible by-products also include all animals that happened to die on
farms or were killed for animal disease eradication, and animals that are not used for human
consumption (zoo and circus animals, pets and fur animals, animals used for experiments
and studies, etc.) .
host, these newly-misfolded prions can transmit the infection. It is not known how normal proteins are
usually able to find the single, correct, folded conformation, among the billions of other possibilities,
without becoming stuck in dead-end intermediates.
10. Daniel Carleton Gajdusek was a Hungarian-Slovak- American medical researcher who was the co-
recipient (with Baruch S. Blumberg) of the Nobel Prize. Gajdusek had the rare distinction of being a
Nobel prizewinner (in Medicine in 1976 for work on kuru) and a convicted child molester. Kuru is an
incurable disease that affects the Fore tribe in Papua New Guinea. Gajdusek showed that it had a long
incubation period, but progresses rapidly when it starts.
1 1 . James Dewey Watson was a biologist, U.S., discovered the structure of the molecule of DNA with
Francis Crick and Maurice Wilkins, with whom he also received the Nobel Prize for Medicine in 1962.
17
Raffaella Leoci
In theory, about 95% of one animal is usable. The remaining 5% are processing losses.
From that 95%, about 55% (on average) of the animal is used for edible products and
the remaining 45% are inedible by-products. According to Meeker and Hamilton 12 ap-
proximately 49% of the live weight of cattle, 44% of the live weight of pigs, 37% of the
live weight of broilers, and 57% of the live weight of most fish species are materials not
consumed by humans. A part of these last by-products are, at times, transformed into
a variety of products used in human food, animal feed, cosmetics, pharmaceuticals and
other technical uses.
Edible animal by-products have traditionally been used as a source of digestible protein,
nutrients, and energy in the feed and animal food industries.
The world production of ABPs derived from the meat and animal production industries
is approximately 60 million tons per year. It has been estimated that more than 10 million
tons of products not destined for direct human consumption, derived from healthy animals,
are produced in the EU every year. During slaughter, between 33 and 43% by weight of the
live animal is removed and discarded as useless waste. A part of these materials, mostly of-
fals 13 are collected and processed by the rendering industry to produce raw materials that are
used in the animal feed industries around the world. 14
Offal, not used directly for human or animal food, 15 is often processed in a rendering
plant producing material that is used for fertilizer 16 or fuel. In some cases, it may be added
to commercially produced pet food.
Typically wastes derive during the processing of ABP and animal products, and more
generally can be identified as occurring from food processing and manufacturing plants,
distribution premises, food markets, wholesale and retail food outlets, and catering facilities
(including household kitchens). On some or any of these sites animal product processing
derivatives include parts of animals deemed unfit for human consumption: butcher and
slaughterhouse waste, catering waste, wool, blood, feathers, hides and skins, used cooking
011. eggs, former foodstuffs, fallen stock, 17 manure, embryos and semen. The by-products
12. Meeker D. L., C. R. Hamilton (2006), An Overview of the Rendering Industry, in Meeker David L.
(Edt), Essential Rendering. All About The Animal By-Products Industry, National Renderers Association,
Printed in September of 2006 by Kirby Lithographic Company, Inc., Arlington, Virginia (USA), p. 2.
13. Offals are the entrails and internal organs (blood, tongue, throat, stomach, brains, honeycomb
tripe, tripe, trepas, tails, sweetbreads, lips, kidney, liver, etc.) of a butchered animal. The word does not
refer to a particular list of organs, but includes most internal organs which are then used in human
food, animal feed, cosmetic, pharmaceutical and technical products.
14. Hamilton C. R. (2002), Real and perceived issues involving animal proteins, Protein Sources For The
Animal Feed Industry, Expert Consultation and Workshop Bangkok, FAO: 29 April - 3 May, 2002.
15- HeartSome offal dishes are considered gourmet food in international cuisine. This includes French
foie gras and pate and sweetbreads. Other offal dishes remain part of traditional regional. This includes
Scottish haggis (sheep stomach stuffed with a boiled mix of liver, heart, lungs, rolled oats and other
ingredients), USA chitterlings (intestines), Jewish chopped liver as well as many other dishes. Intestines
are used as casing for meat sausages or blood sausage (boudin noir, or the Spanish "morcilla", or Ital-
ian "sanguinaccio") For further notices see: Helou A. (2004), Offal, the Fifth Quarter, Absolute Press,
Scarborough House, Bath, UK (last edition 201 1).
16. Sharrock P., M. Fiallo, A. Nzihou, M. Chkir (2009), "Hazardous animal waste carcasses transfor-
mation into slow release fertilizers" , Journal of Hazardous Materials, vol. 167 (1-3): 1 19-123.
17. "Fallen stock" can be any animal that has died of natural causes or disease on the farm or has been
killed on the farm for reasons other than human consumption.
18
Animal by-products (ABPs): origins, uses, and European regulations
are destined for destruction because they are not usable in the consumer market or to the
fertilizer or the livestock industries, nor are they suitable for the production of biogas or for
composting. In other words, if no part of the ABPs is separated and used, the ABPs, on the
whole, are considered waste that must be disposed.
ABPs and wastes are a potential source of risks to public and animal health. Improper use
of ABP has resulted in outbreaks of serious diseases such as bovine spongiform encephalopathy,
and as already said, foot and mouth disease, avian influenza, classical swine fever, etc.
3. Historical uses of ABPs
The process of rendering animal parts has been documented for more 2,000 years. 18 The
purpose of rendering was to produce tallow and other animal fats to make candles and soap.
In the past, non-food, non-feed applications for rendered ABPs, with the exception of
tallow, were limited in their application to niche markets 19 that were too small to support
large volumes of meat and bone meal.
Nowadays, while muscles are the more consumed part of the animals, ABPs such as the
entrails and internal organs are also widely consumed. The use and value of inedible meat
by-products and offal-edible, considered waste material to be thrown away or high-priced
delicacies, depend on the country in question and its culture. The blood, bones, meat trim-
mings, skin, fatty tissues, glands, horns, hoofs, lags and feet, and skulls of harvested ani-
mals, comprise a wide variety of products. When slaughtering healthy animals and collecting
slaughterhouse debris, those natural and environmentally compatible by-products become
raw materials for the industry in general. 20
Historically, non-feed, non-food applications for ABPs, excluding tallow, has tended to
be limited in their application to niche markets. 21 Historically, "rendering" has been defined
as separation of fat from animal tissues by the application of heat. Rendering can refer to any
processing of animal products into more useful materials, or more narrowly to the rendering
of whole animal fatty tissue into purified fats like tallow or lard. Rendering can be carried
out on an industrial, farm, or kitchen scale.
Meeker and Hamilton 22 noted that "one-third to one-half of each animal produced for
meat, milk, eggs, and fiber is not consumed by humans. These raw materials are subjected
to rendering processes resulting in multiple useful products. Meat and bone meal, meat
meal, poultry meal, hydrolyzed feather meal, blood meal, fish meal, and animal fats are the
primary products resulting from the rendering process". The same authors stated, "A by-
product is defined as a secondary product obtained during the manufacture of a principal
commodity. A co-product is a product that is usually manufactured together or sequentially
18. Grummer R. R. (1992), Inedible Fats and Greases, Chapter 6, Edts., A. E. Pearson and T. R. Dut-
son, Inedible Meat By-Products, Elsevier Applied Science, London-New York, pp. 113-148.
19. Pearl G. G. (2003), Non-Food/Feed Uses of Rendering Products: Identification of New Opportu-
nities and Assessment of Major Barriers to their Exploitation, Fats and Protein Research Foundation,
Research Project #01B-6, Bloomington, Illinois (USA), pp. 4-35.
20. Ockerman H., C. L. Hansen (2000), Animal by-product processing & utilization, Technomic Pub-
lishing Company Inc., Lancaster (PA- USA).
21. Pearl G. G. (2003), "Non-feed, non-food applications for animal by-products", Render, 32 (1):
22-25.
22. Meeker D. L., C. R. Hamilton, An Overview of the Rendering Industry, ibid., p. 1 .
19
Raffaella Leoci
with another item because of product or process similarities". We will preferably use the
term by-product. Rendering is a process of both physical and chemical transformation using
a variety of equipment and processes. All of the rendering processes involve the application
of heat, the extraction of moisture, and the separation of fat.
According to Romans and others, 23 rendering involves the heating or cooking of raw
materials, with complex or simple mixtures of protein, minerals, and fatty substances, to
liquefy fats and break down membranes or other structures that may hold fat. According to
Kumar, 24 the goals of carcass rendering are elimination of water, separation of fat from other
materials (mainly protein substances), sterilization of the final products, and production of
MBM 25 from a variety of condemned, fallen, culled, and experimental animals.
Prokop, 26 UKDEFRA, 27 Romans and others defined rendering as a process of using
high temperature and pressure to convert whole animal and poultry carcasses or their by-
products with no or very low value to safe, nutritional, and economically valuable products.
In fact, the highly perishable protein and fat materials comprising carcasses become a major
problem and a liability if they are not converted, stabilized, or somehow processed during
the 24 hours following death.
A lot of industrial chemical applications have used fat and fatty acids as a feedstock,
whereas relatively few applications were developed for ABPs beyond adhesives and fertilizers.
The outbreak of World War I and II saw great demand for glycerin used in the production of
explosives (tri- nitroglycerin). The demand for recovering co-products for these applications
has declined with the increase in use of widely available petroleum/synthetic-based products.
In order to add value to animal by-products of the hide and internal organs, the meat in-
dustry is using science and innovation. 28 Industry is using science and research to add value
to ABPs far beyond its usual profitability. It is still necessary to employ the most up-to-date
and effective research tools to study these products for their nutritional properties, to search
for medical, cosmetic and in other industrial fields, to develop new technological applica-
tions and to continue innovation towards advanced value-addition of ABPs.
23. Romans J. R., W. J. Costello, C.W. Carlson, M. L. Greaser, K. W. Jones (2001), Packing house
by-products, in The Meat We Eat, Interstate Publishers, Inc., Danville, Illinois; Jimenez-Colmenero F.,
M. Reig, F. Toldra (2006), New Approaches for the Development of Functional Meat Products, Chapter
1 1 , in L. M . L . Nollet and F. Toldra (Edtrs) , Advanced Technologies For Meat Processing, CRC Press,
Florida (USA), pp. 275-308.
24. Kumar M. (1989), Handbook of rural technology for the processing of animal by-products (FAO Agri-
cultural Services Bulletin No. 79), FAO of the United Nations, Rome.
25. MBM (Meat and Bone Meal) is prepared from the rendering of dead animals or wastes materials
associated with slaughtering operations (carcass trimmings, condemned carcasses, condemned livers,
inedible offal and bones). It is basically dry rendered protein product from mammal tissues with more
than 4.4% phosphorus.
26. Prokop W. H. (1996), The rendering industry — a commitment to public service, in D.A. Franco &
W. Swanson (Edts.), The original recyclers, Joint publishers: the Animal Protein Producers Industry, the
Fats & Proteins Research Foundation, and the National Renderers Association, Merrifield, VA (USA).
27. UKDEFRA (2000), The BSE inquiry report, Vol. 13: Industry processes and controls, Ch. 6 render-
ing, Annex B: manufacturing processes of rendering.
(See: http://www.bseinquiry.gov.uk/report/volumel3/chapterj.htm).
28. See: Toldra E, M. Conception Aristoy, M. Reig (2012), "Innovations in value-addition of edible
meat by-products", Meat Science, 92 (3): 290-296.
20
Animal by-products (ABPs): origins, uses, and European regulations
As noted by Peter Kent in Clemson Impacts: 2 ' "The new Clemson Animal Co-Products
Research and Education Centre was dedicated in March (2006). This research initiative will
serve the rendering industry, which collects and processes billions of pounds of animal remains,
the inedible leftovers from meat production. Clemson researchers will seek new ways to recycle
fats and protein from food animal production to create a variety of value-added products, in-
cluding bio-fuels, fertilizers, soap, rubber, and plastics. Also, the center will work to ensure the
safety of rendered products for animal feeds and consumer products, promote environmentally
sound practices, and provide educational opportunities in utilizing animal co-products".
McGlashan, 30 in his preamble for the Clemson University Animal Co-Products Re-
search and Education Center dedication conference (April 2006), states: "It is imperative
to society that the rendering industry remains viable". As stated in the books first chapter,
"Overview of the Rendering Industry", the availability of rendered products for animal feeds
in the future depends on regulation and the market. Future regulation relating to biosecu-
rity and environmental protection has the potential to restrict traditional market access for
rendered co-products. Hence, it is essential that new applications and avenues for profit-
able disposal of co-products are discovered, researched, developed into a viable commercial
process, and widely adopted by the industry in order to maintain rendering as a viable and
valuable service to the meat processing sector".
Researchers at many universities believe that these inedible parts of carcass may have
value. Researchers launched their research initiative after finding that there is a lack of cen-
tral location for expertise in determining value from animal agriculture that is not simply
muscle related. For example, Schaefer 31 stated: The mission "is really to discover new uses
of known bioactive molecules, mass produced from by-products of animal and poultry that
have been produced for meat, and to discover novel compounds that can be used in biology,
agriculture, medicine and other consumer products".
The UKDEFRA 27 recognized that animal waste collection and rendering "constituted
a vital public service as well as commercial activity" but made some recommendations in-
tended to remedy the effect on competition of these firms' pricing policies.
4. Current uses of ABPs
At present, the main ABPs useful for foods, industrial products and pharmaceutical
uses, are the following:
• Bile: detergent, pharmaceuticals;
• Blood:
a. Edible blood: additives, binders or meat glue, blood flours, blood proteins, blood
sausages, cake mixes, clarifiers, coffee whiteners, curing agents, deep-fry batters,
dyes and color enhancers, egg albumin substitutes, emulsifiers, fat replacer, gravy
mixes, imitation seafood, pasta, stabilizers, whipped toppings;
29. Clemson Impacts, a quarterly publication of Clemson Public Service Activities, is available on the
web www.clemson.edu/public/.
30. McGlashan S. A. (2006), Industrial And Energy Uses Of Animal By-Products, Past And Future, con-
ference held at the Madren Conference Center on campus in March 27-29, 2006, p. 242.
31. D. M. Schaefer, Professor and Department Chair, Department of Animal Science, 256A Animal
Science Bldg, 1675 Observatory Drive, Madison, WI 53706.
21
Raffaella Leoci
b. Feed (blood meal): lysine supplement, milk substitute, nutritional component, and
vitamin stabilizer;
c. Fertilizer: mineral components, seed coating, soil pH stabilizer;
d. Scientific use or research: albumin, antibodies, antiserum, blood clotting factors,
leukocytes - white blood cells, plasma, red blood cells, serum, whole blood, antigens;
e. Laboratory uses: serum, culture media, tannin analysis, active carbon, haemin,
blood agar, fibrinogen, peptone, glycerophosphates, coagulation factors, blood
albumin, RH factor typing, globulins, sphingomyelins, catalase, agglutination tests,
diagnostic microbiology;
f. Pharmaceutical use: immunoglobulin, thrombin (blood clotting), factors, fibrinogen,
fibrinolysin, fibrin products, serotonin, kallikreins, plasminogen, plasma extenders,
transfusion. Products usually for manufacturing cosmetics, human nutritional
supplements, and test kit components;
g. Veterinary biological use (also called veterinary biologies);
h. Industrial uses: glue and resin extender, finishes for leather and textiles, insecticide
spray adjuvants, egg albumin substitute, foam fire extinguisher, porous concrete,
ceramic and plastic manufacture, plastic and cosmetic base formulations.
• Bones: adhesives, animal feed, bandage strips, bone meal (calcium and phosphorous
source), bone marrow (blood disorders), cold cream, collagen and bone for plastic sur-
gery, crochet needles, cutlery handlers, dice, dog biscuits, emery boards and cloth,
glycerin, fertilizer, glue, gelatin, hairpins, imitation ivory, inedible bone meal, livestock
feeds, meal, mineral source in supplements, neat fat oil, piano keys, photographic film,
plant food, plywood and paneling, protein hydrolysate, syringes, soft cartilage (plastic
surgery), shampoo and conditioner, tallow and ornaments, wallpaper and wallpaper
paste, xiphisternal cartilage (breastbone);
• Brains and spinal cords: steroid, cholesterol, lecithin, cephalin;
• Fats and fatty acids: antifreeze, animal foods, biodegradable detergents, biodiesel, cello-
phane, cement, ceramics, chalk, chemicals, cosmetics, crayons, creams and lotions
(sheep), deodorants, detergents, explosives, fertilizer, fiber softeners, floor wax,
glycerin, glycerol, herbicides, horse and livestock feeds, industrial oils, insecticides,
insulation, linoleum, livestock feed, lubricants, lubricating greases, makeup, matches,
medicines, mink oil, nitroglycerine, oil polishes, ointment bases, oleostearin, paints,
paraffin, perfumes, pet foods, pharmaceuticals, plasticizers, plastics, printing rollers,
protein hair conditioner, protein hair shampoo, putty, rubber products, shaving cream,
shoe cream, soaps, solvents, stearic acid (sheep), tallow for tanning, textiles, tires, water
proofing agents, weed killers;
• Glands:
ft. Adrenal: cortisone (for arthritis, skin allergies, anti-inflammatory
medicine), epinephrine (aid in raising blood pressure, heart disorders,
and allergies);
b. Kidney: pharmaceuticals and others (for transplantation);
c. Liver: liver tonics, heparin (anti-coagulant, prevents gangrene), liver extract (treat-
22
Animal by-products (ABPs): origins, uses, and European regulations
ment of anemia), intrinsic factor (pernicious anemia), Vitamin B12 (prevention of
B-complex deficiencies);
d. Pancreas: chymotrypsin (contact surgery), diastase (aid in starch digestion), gluca-
gon (treat hypoglycemia), insulin (diabetes mellitus), pancreatin (aid digestion),
trypsin (for burns, wounds, and infection, promotes healing, aid in protein diges-
tion and in cleaning wounds);
e. Pituitary glands: ACTH (arthritis, allergies, rheumatic fever, skin and eye inflam-
mations), pressor hormone (regulates blood pressure), prolactin (promotes lacta-
tion), vasopressin (controls intestinal and renal functions);
f. Spleen: in addition to food, ferritin, for stimulating the immune system;
g. Thymus: thymosin, thymopoeitin, thymulin, thymic humoral factor, thymus
extract, as food;
h. Thyroid (Thyrar): TSH (thyroid diagnosis thyroid extract - hypothyroidism,
thyroid hormones, myxedema cretinism.
• Hides and skins: bandages, belts, bookbinding, cabinetmaking, collagen-based
adhesives (from trimmings), drum head (sheep), emery boards, gelatin, gloves, and shoes,
glues-for papermaking, leather, leather sporting goods, leather wearing apparel,
luggage, pharmaceuticals, photographic materials, pigskin garments, porcine burn
dressings for burn victims, sheetrock, shoes and boots, upholstery, wallets, wallpaper;
• Hairs and wools, skins, feathers, nails, horns, and hooves: air filters, artist's paint brush,
brushes, fabrics, feather meal, crochet needles, hairpins, imitation ivory, horn handles,
cutlery handlers, buttons, cellophane wrap and tape, laminated wood products, chess-
men, combs, protein hydrolysate, fibers, felt and rug padding, insulation material,
non-woven, plastering material, textiles, upholstering material, yarn;
• Hearts (hog heart valves for human transplant);
• Intestines: sausage casing, strings (for suturing material, surgical ligatures, musical
instruments, racquets), heparin. Small Intestine Submucosa (SIS materials);
• Lungs: heparin (anti-coagulant, prevents gangrene), meal;
• Meat and bone meals (pyrolysis, fluidized bed reactor, anaerobic digestion, co-firing/
incineration, concrete and asphalt construction);
• Ovaries: estrogen (progesterone, relaxin, etc.);
• Proteins: protein hydrolysis, plastics, adhesives, surfactant;
• Stomach and tripe: pepsin (aid in protein digestion), rennet (aid in milk digestion);
• Trachea: chondroitin sulphate.
As can be seen, some products can be obtained from various components of the ABPs,
for example glues and gelatins. Blood, bones, skins, horns, feathers, as we will see later, can
be used for production of glues, while bones, skin and connective tissue are used for the
production of gelatin which is then used in human food, animal feed (coating on vitamins,
and binders on feed pellet and dog chews), pharmaceutical manufacturing (hard and soft
capsules) and technical use (in the photographic industry, for example, as paper coating and
as a component in silver halide emulsion coatings, etc.). Gelatin is the main ingredient in
23
Raffaella Leoci
products like Jell-O and Gummy Bears and can be found in many other product, such as
yogurt. Gelatin is usually made by boiling the skin, tendons, ligaments, and bones of cows
and pigs after the meat has been harvested. It can also be made from fish or other animals.
The alternative to gelatin is "agar- agar" 32 which is produced from seaweed.
Fertilizer and soil conditioners are a minor factor for ABPs while there is widespread and
profitable use of protein-based meals in the feed and food sector and valuable use of many
components of ABPs in different industrial sectors.
The products for general food are the following:
1 . from cattle, sheep, hog flesh: a great variety of fresh, frozen, and pre-cooked meats and
prepared and processed meat products;
2. from milk/ dairy: butter, casein, cheese and cheese products, cream, food ethanol, ice
cream and ice cream mixes, lactose (carbohydrates), milk powder, sherbet, whey (pro-
teins), fats (lipids), yogurt;
3. from fats and fatty acids: chewing gum, lard, oleo margarine, oleo shortening, oleostea-
rin, pharmaceuticals, rennet for cheese (sheep), rennet for cheese (sheep), shortening.
Some of the above by-products contain highly valued compounds. The high market
price of these compounds/products provide profitable opportunity for the "mining" of lipo-
proteins, peptides, enzymes, hormones, insulin, etc. that can be extracted from glands and
organs, concentrated, dried for storage, and further processed or sold as raw material for
other industries.
The likely future use of organs (hearts, livers, intestines, lungs, etc.) taken from ge-
netically modified animals may become objects of primary production, while the meat from
these animals could assume the role of ABP, if not intended for human consumption.
Premises handling ABPs include:
• rendering plants;
• handling and storage plants (previously known as intermediate plants);
• incinerators/co-incineration plants;
• composting and anaerobic digestion (biogas) plants;
• pet food plants;
• technical plants;
• collection and treatment plants for ABP;
• final users of treated ABP;
• pet food plants;
• rendering plant.
Let us take a closer look at the most important uses of above mentioned ABPs.
32. Agar-agar is a polysaccharide that comes from Japan used as a gelling natural. It derives from red
algae belonging to different genera (Gelidium, Gracilaria, etc.). It is a polymer consisting of units of
D-galactose. The galactose is one of the two components of the lactose, that is a disaccharide formed
from one molecule of a-glucose and of a (3-galactose.
24
Animal by-products (ABPs): origins, uses, and European regulations
4.1 Bile
Bile is a digestive fluid that is made and released by the liver and stored in the gall-
bladder. Bile helps break down fats into fatty acids, which can be taken into the body by
the digestive tract. Bile contains mostly cholesterol, bile acids (also called bile salts), and
bilirubin (a breakdown product of red blood cells). It also contains water and body salts
(potassium and sodium), as well as very small amounts of copper and other metals.
The first studies on the use of bile salts as detergents (anionic detergents) date back to
the years prior to World War II. 33 It was observed at the time, but the characteristic was
already known, 34 as "the synthetic detergents and the bile salts all have the same general type
of hydrophobic-hydrophilic structure. Each consists of a large hydrophobic part with a small
hydrophilic part attached to it". Bile powder is used as nutraceutical for the treatment of
indigestion, for constipation and for bile tract disorders. It is also used to increase the secre-
tory activity of the liver and in aquaculture as a prawn feed ingredient. Mixed bile acids are
also used in poultry and pig foods. Natural taurine 35 is used in infant milk formula to make
it more like human milk, in health tonics ("smart drinks"), and for the prevention of side
effects from excess alcohol consumption.
Gallstones 36 are reported to have aphrodisiac properties and can be sold at a high price.
They are usually used as ornaments to make necklaces and pendants.
As pharmaceutical use, bile acids or their salts and conjugates 37 are formulated for non-
steroidal anti-inflammatory drugs, such as indomethacin, 38 and other pharmaceutically ac-
tive compounds (antigen-vaccine preparation, cell culture, core bioreagents, etc.). Bile is
manufactured from raw ox bile and contains a mixture of conjugated bile acids, namely a
mix of bile salts, predominantly sodium glycocholate hydrate (C 26 H 42 NO s Na-nH 2 0) and
sodium taurocholate hydrate (C 26 H 44 NNa0 7 S-nH 2 0) along with a small amount of free bile
acids (cholic acid) and lipids. The bile salts are used as an ingredient in the manufacturing
of dehydrated culture media, such as McConkey, VRBL, Electrophoresis Chromatography,
and others. A major use is in clinical microbiology to selectively grow fecal staphylococci and
streptococci (MacConkey Agar).
33. Anson M. L. (1939), "The denaturation of proteins by synthetic detergents and bile salts", The
Journal of General Physiology, 22: 239-246.
34. See: Kuhne W., L. Hermann (1879), Handbuch der Physiologie, EC.W. Vogel, Leipsic, p. 264.
35. Taurine (C 2 H ? N0 3 S) is present in most animal tissues and is particularly abundant in oysters,
mussels, bovine bile and human breast milk. It is one of the amino acids classified as non-essential,
which means it can be made by the human body. Research also suggests that it might also protect cell
membranes from toxic substances. In fish it is thought to be an osmoregulator of cell volume.
36. Gallstones are hard, pebble-like deposits that form inside the gallbladder.
37. See: Barnwell S. G., Pharmaceutical composition containing a low detergent effect bile salt and
an active compound that undergoes biliary excretion and/or enterohepatic recycling, US Patent
5,942,248, 1999.
38. Indomethacin is a nonsteroidal anti-inflammatory drug that reduces fever, pain and inflammation.
It is similar to ibuprofen and naproxen.
25
Raffaella Leoci
Ursodeoxycholic acid 39 is found naturally in bear bile 40 . It is an Asian folk medicine
used to protect the liver. Modern medical research has proved that ursodeoxycholic acid is
an effective drug in the treatment of liver disease. 41 Cholic acid (C 24 H 40 O 5 ) 42 is used as the
starting material in the synthesis of ursodeoxycholic acid.
Cholic and deoxycholic acids are the starting point for the synthesis of many steroids.
They are used as starting materials for other steroidal compounds such as gallstone-dissolv-
ing drugs, hepatic drugs, digestive aids, and corticosteroids.
Bile powder is used as nutraceutical to aid digestion and in aquaculture as a prawn feed
ingredient. Mixed bile acids are also used in prawn feed and in poultry and pig foods also.
Natural taurine (an amino acid, not a steroid) is used in infant milk formula to make
it more like human milk, in health tonics ("smart drinks"), and for the prevention of side
effects from excess alcohol consumption.
It is used some ingredients of bile, such as prednisone and cortisone, can be extracted
separately, and used as medicines.
4.2 Blood
Blood or blood fractions are derived from ruminants, birds or poultry, equine (horses),
and swine.
Blood, as it is known, is composed of blood cells (40% of total blood volume) sus-
pended in a liquid blood plasma. Its composition is 18-19% protein (fibrinogen) and 78-
79% moisture. Blood cells are composed of red cell (erythrocyte), white cell (leukocytes),
and platelets (blood clotting). Plasma contains globulins, albumins, fibrinogen, dissipated
proteins, glucose, mineral ions, hormones, carbon dioxide, and blood cells themselves.
The color of beef plasma is yellow or orange and that of pig plasma is gray-white to pink.
Blood plasma proteins, such as albumin and globulins, are good emulsifiers.
Animal blood amounts to 5-7% of the body weight of mammals (dried blood makes
up to 0.7% of live weight). While half of the blood volume of a slaughtered animal remains
in the carcass tissues (50% retained in the capillary systems throughout the body) and is
eaten with the meat and internal organs, the other half recovered from bleeding represents
5-8 percent of the protein yield of a slaughtered animal. The remainder (42-45%), if it is
not recovered may cause environmental pollution hazard. In the future, we cannot afford to
waste such large amounts of animal protein.
39. Ursodeoxycholic acid is one of the secondary bile acids, which are metabolic by-products of intes-
tinal bacteria. The liver produces primary bile acids and stores it in the gall bladder. Primary bile acids
when secreted into the intestine can be metabolized into secondary bile acids by intestinal bacteria.
Primary and secondary bile acids help the body digest fats. Ursodeoxycholic acid helps regulate cho-
lesterol by reducing the rate at which the intestine absorbs cholesterol molecules while breaking up
micelles containing cholesterol. Because of this property, ursodeoxycholic acid is used to treat (choles-
terol) gallstones non-surgically.
40. Bears suffer terribly when people extract the bile for medicinal use. Bears cannot be used as a source
where large quantities of it are required as a pharmaceutical.
41. Paumgartner G., U. Beuers (2002), "Ursodeoxycholic Acid in Cholestatic Liver Disease: Mecha-
nisms of Action and Therapeutic Use Revisited", Hepatology, 36 (3): 525-31.
42. One of the primary bile acids, usually found conjugated with glycine or taurine. It facilitates fat
absorption and cholesterol excretion.
26
Animal by-products (ABPs): origins, uses, and European regulations
Blood used for transfusions is always human in origin, though some blood substitutes are
made from animal sources. Many diagnostic laboratory tests use animal or human sourced
reagents. In recent years transfusion medicine has become very prevalent in veterinary care.
Animal blood banks do supply blood products.
The procedures for edible blood collection undergo different phases. Healthy animals
must be inspected and passed for use as meat food. After collection, blood needs to pass in-
spection. Then it can be used for human utilization since blood taken from healthy animals
is essentially sterile and any contamination is due to the bleeding technique and the drain-
age system employed during collection. 43 Many blood products are made from the plasma
component of blood.
Let's take a look at the main uses.
a) Edible blood (Food)
The meat industries use the blood plasma and isolated blood proteins as ingredients in the
food industry, mainly as a binder but also as natural color enhancers, emulsifiers, fat replacers,
meat curing agents, etc. Blood plasma and blood proteins function as a binder in meat systems
thanks to their ability to form gels upon heating. Other industries use blood albumin, meat
glue and the bulk of the blood in blood sausage, soup, and as protein supplement. 44
Additives
Blood is an abundant source of iron and proteins of high nutritional and functional
quality. We will see below the use of proteins for alimentary use. Extensive studies and pat-
ent searches, on the functional properties of blood, have shown that blood plasma exhibits
many functional properties, including, gelling, water holding, solubility, emulsification and
foaming capacity.
Plasma is widely used in the food industry because it possesses a neutral taste and is
devoid of the dark color associated with red blood cells. Instead, hemoglobin is utilized for
its color. The uses are as follows:
• Porcine or bovine plasma, with increased fibrinogen concentration, is used as cold
set binder for meat products;
• Bovine spray-dried plasma concentrate as emulsifier, gelling and binding agent in
meat and fish-based products, pasta and bakery products;
• Porcine or bovine stabilized hemoglobin, porcine or bovine frozen or powder hemo-
globin as natural coloring for meat products;
• Porcine or bovine natural colorant obtained from the red pigments of blood in order
to enhance meat color and increase contrast between fat and meat;
• Bovine globin (hemoglobin with the heme group removed) as emulsifier in meat products;
• Porcine or bovine heme iron polypeptide for iron supplementation.
Blood and its other derivatives, has the potential to serve as a useful source of valuable
peptides with antioxidant, anti-hypertensive, and anti-bacterial properties. Several bioactive
43. Riaz M. N. (2010), "Fundamentals of halal foods and certification", Prepared foods, 179 (1): 71-76.
44. The proteins content, of high quality, is about 18% of the blood (see: Ockerman H. W., C. L.
Hansen (2000), ibid., pp. 325-353).
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Raffaella Leoci
peptides that offer other health benefits such as analgesic and antinociception activities, have
also been isolated from blood proteins. 45
Iron is a mineral that is necessary for producing red blood cells and for redox processes. 46
Iron deficiency is the most common nutritional deficiency in the world since it affects about
20% of the world's population, with children and women at greater risk. 47 The primary
cause of iron deficiency, apart from iron loss caused by blood loss through parasite infesta-
tion (for example, hookworm), is the inability to satisfy iron requirements with dietary
iron 48 intake. In the poorer regions of the world (especially Asia and Africa) iron is provided
mainly by plant-based diets that usually contain non-heme iron, a form of iron that is poorly
absorbed. 45 Lack of iron in diets can cause many problems, may lead to unusual tiredness,
shortness of breath, a decrease in physical performance, 50 and learning problems in children
and adults, and may increase probability of getting an infection. In adults, low iron levels
impair the ability to do physical work. 51 Iron deficiency, on the other hand, can proceed to
iron-deficiency anemia and other adverse conditions, including at-risk pregnancy outcomes
and, possibly, impaired intellectual development in infancy. 52
Blood is a good source of bioavailable iron. As is well known, cooking decreases the
bioavailability of iron and its absorption is improved when delivered in a meat-based diet:
during digestion of meat, ferric iron can be converted into the bioavailable ferrous form.
45. See: Teschemacher H. (2003), "Opioid Receptor Ligands Derived from Food Proteins", Current
Pharmaceutical Design, 9 (16): 1331-1344; Gomes I., C. S. Dale, K. Casten, M. A. Geigner, F. C.
Gozzo, E. S. Ferro, A. S. Heimann, L. A. Devi (2010), "Hemoglobin-derived peptides as novel type of
bioactive signalling molecules", AAPS Journal, 12 (4): 658-669; CoderreT. J., I. Van Empela (1994),
"The utility of excitatory amino acid (EAA) antagonists as analgesic agents. I. Comparison of the
antinociceptive activity of various classes of EAA antagonists in mechanical, thermal and chemical
nociceptive tests", Pain, 59 (3): 345-352; Aslaniants Zh. K., G. R. Melik-Eganov, A. V. Evstratov,
M. P. Ivanov, S. G. Batrakov, N. V. Korobov, V. V. Iasnetsov (1991), "The effect of blood serum pro-
teins from the seal on the analgetic action of narcotic analgesics", Bulleten Eksperimentalnoi Biologii i
Meditsiny (Moskva), 112 (11): 503-5.
46. Lieu P. T, M. Heiskala, P. A. Peterson, Y. Yang (2001), "The roles of iron in health and disease",
Molecular Aspects of Medicine, 22 (1-2): 1-87.
47. Assessing the iron status of populations. Report of a Joint World Health Organization, Centers
for Disease Control and Prevention, Technical Consultation on the Assessment of Iron Status at the
Population Level, 6-8 April 2004, Geneva, Switzerland; Martmez-Navarrete N., M. M. Camacho, J
Martmez-Lahuerta, J. Martmez-Monzo, P. Fito (2002), "Iron deficiency and iron fortified foods - a
review", Food Research International, 35 (2-3): 225—231.
48. Hoppe M., L. Hulthen, L. Hallberg (2008), "The importance of bioavailability of dietary iron in rela-
tion to the expected effect from iron fortification", European Journal of Clinical Nutrition, 62 (6): 761-769.
49. Chiplonkar S. A, K. V. Tarwadi, R. B. Kavedia, S. S. Mengale, K. M. Paknikar, V. V. Agte (1999),
"Fortification of vegetarian diets for increasing bioavailable iron density using green leafy vegetables",
Food Research International, 32 (3): 169-174; Yi-Chia Huang (2000), "Nutrient intakes and iron status
of vegetarians", Nutrition, 16 (2): 147-148.
50. Gardner G. W., V. R. Edgerton, B. Senewiratne, R. J. Barnard, Y. Ohira (1977), "Physical work
capacity and metabolic stress in subjects with iron deficiency anemia", The American Journal of Clinical
Nutrition, 30 (6): 910-917.
51. Haas J. D., T. Brownlie (2001), "Iron deficiency and reduced work capacity: a critical review of the
research to determine a causal relationship", Journal of Nutrition, 131 (2S-2): 676S-690S.
52. Baynes R. (1990), "Iron Deficiency", Annual Review of 'Nutrition, 10 (1): 133-148.
28
Animal by-products (ABPs): origins, uses, and European regulations
Several strategies are commonly used to combat iron deficiency, including supplementa-
tion with capsules and tablets and fortification of processed foods. 53 However, in developing
countries, food-based strategies remain the most sustainable approach for addressing iron
and other micronutrient deficiencies. 54
Fortifying staple foods with heme-iron, which is better absorbed than non-heme iron 55
because its absorption is essentially unaffected by other dietary factors, has therefore been
suggested as a measure to overcome the problem of iron deficiency.
As bovine blood has the largest amount of heme iron than any animal source and is
readily available, it has been suggested for the fortification of staple foods. Hertrampf and
others 56 field tested an extruded rice flour, fortified with a bovine hemoglobin concentrate
(Fe:l4 mg/100 g of powder). Their results demonstrate that the consumption of a hemo-
globin-fortified cereal is effective in markedly reducing the incidence of iron deficiency in
breast-fed infants. Walter and others, 57 investigated the effect of bovine-hemoglobin-fortified
cookies on the iron status of school children in a nationwide school lunch program in Chile.
According to Haschke and others 58 feeding infants iron-fortified (12-15 mg/1) formulas is an
effective and convenient means to protect infants from iron deficiency. Quintero-Gutierrez
and others 59 evaluated the bioavailability of heme iron added to biscuit filling. It comprised
two stages: first, the development of the heme iron enriched biscuit filling, second, the evalu-
ation of the bioavailability of the mineral in fattening piglets. The heme iron was prepared
53. Suparat Reungmaneepaitoon, Chomdao Sikkhamondhol, Chansuda Jariyavattanavijit, Chow-
ladda Teangpook (2008), "Development of instant noodles from high-iron rice and iron-fortified rice
flour", Songklanakarin Journal of Science and Technology, 30 (6): 713-721.
54. FAO and ILSI, Preventing micronutrient malnutrition a guide to food-based approaches - Why
policy makers should give priority to food-based strategies, Washington, DC 1997.
55. The chemical state of iron are ferrous (*Fe 2 ) that is more easily absorbed than ferric ( + Fe 3 ) and
this change is largely dependent on the presence of luminal gastric acid and ceruloplasmin within the
intestinal mucosal cell. Heme and non-heme are two forms of dietary iron. The difference between
heme and non-heme iron is that the first is derived from haemoglobin while non-heme irons do not
come from an animal source. Since the iron in animal-based foods is about 40% heme iron and 60%
non-heme iron, animal-based foods are good sources of absorbable iron (See: Hallberg L., L. Hulthen
(2000), "Prediction of dietary iron absorption: an algorithm for calculating absorption and bioavail-
ability of dietary iron", The American Journal of Clinical Nutrition, 71 (5): 1147-60.
56. Hertrampf E., M. Olivares, F Pizarro, T. Walter, M. Cayazzo, G. Heresi, S. Llaguno, P. Chadud, A.
Stekel (1990), "Haemoglobin fortified cereal: a source of available iron in breast fed infants", European
Journal of Clinical Nutrition, 44 (11): 793-798; Kikafunda J. K., P. Sserumaga (2005), "Production
and use of a shelf-stable bovine blood powder for food fortication as a food-based strategy to combat
iron deficiency anaemia in Sub-Saharan Africa", African Journal of Food Agriculture Nutrition and De-
velopment, 5 (1): 1-18.
57. Walter T, E. Hertrampf, F. Pizarro, M. Olivares, S. Llaguno, A. Letelier, V. Vega, A. Stekel (1993),
"Effect of Bovine-Hemoglobin Fortified Cookies on Iron Status of Schoolchildren - a Nationwide
Program in Chile", The American Journal of Clinical Nutrition, 57 (2): 190-194.
58. Haschke E, H. Vanura, C. Male, G. Owen, B. Pietschnig, E. Schuster, E. Krobath, C. Huemer
(1993), "Iron nutrition and growth of breast- and formula-fed infants during the first 9 months of
life", Journal of 'Pediatric Gastroenterology and Nutrition, 16 (2): 151-6.
59. Quintero-Gutierrez A. C, G. Gonzalez-Rosendo, J. Sanchez-Munoz, J. Polo-Pozo, J. J. Rodriguez-
Jerez (2008), "Bioavailability of heme iron in biscuit filling using piglets as an animal model for hu-
mans", International Journal of Biological Science,. 4 (1): 58-62.
29
Raffaella Leoci
by isolating hemoglobin from porcine blood followed by enzyme hydrolysis and subsequent
separation of globin from the heme group using ultrafiltration. They showed that an accept-
able product with high heme iron content can be formulated and suitable for use as biscuit
filling. The heme iron supplement produced weight increase and a lessened mortality in
fattening pigs. A similar study was conducted by Gonzalez-Rosendo and others. 60 They used
a heme-iron concentrate product derived from swine hemoglobin to enrich the chocolate-
flavored filling of biscuits and the bioavailability of this source of heme-iron was assessed in
adolescent girls. The study showed that the iron contained in the heme-iron concentrate was
well absorbed and tolerated by the adolescents included in the study. Similarly Olivares and
others support the study 61 . During their research school-age children in Chile received 30 g
of wheat-flour biscuits daily through a "National School Lunch Program". To improve iron
nutrition, the biscuits were fortified with a 6% bovine hemoglobin concentrate. The high-
iron bioavailability, the good organoleptic characteristics and the biological effect on iron
nutritive made the product an appealing alternative to combat iron deficiency.
However, some 62 believe that it is impractical to use hemoglobin as an iron supplement
because it is low in iron. Vaghefi and others 63 assert at the end of their study that heme
iron absorption depends not only on its solubility but also relies mainly on the balance
between the strength of heme-peptides and the polymerization rate of heme. Others 64 state
that heme-iron is poorly soluble at low gastric pH, and its absorption would be less than
iron absorption from muscle. 65 Hallberg and others 66 observe that since Ca inhibits the ab-
sorption of heme- and non-heme-Fe to the same extent, their results strongly suggest that
Ca interferes with the transport of Fe through the mucosal cell and, at a late stage, is com-
mon for heme- and non-heme-Fe transport. They conclude that observations of Ca strong
60. Gonzalez-Rosendo G., J. Polo, J. J. Rodriguez-Jerez, R. Puga-Di'az, E. G. Reyes-Navarrete, A. G.
Quintero-Gutierrez (2010), "Bioavailability of a heme-iron concentrate product added to chocolate bis-
cuit filling in adolescent girls living in a rural area of Mexico" ', Journal of Food Science, 75 (3): 73H-78H.
61. Olivares M., E. Hertrampf, E Pizzarro, T. Walter, M. Cayazzo, S. Llaguno, P. Chadud, N. Carta-
gena, V. Vega, M. Amar, A. Stekel (1990), "Hemoglobin-fortified biscuits: bioavailability and its effect
on iron nutriture in school children", Archivos latinoamericanos de nutrition, 40 (2): 209-220.
62. For review see: West A. R., P. S. Oates (2008), "Mechanisms of heme iron absorption: Current
questions and controversies", World Journal of Gastroenterology, 14 (26): 4101-4110.
63. Vaghefi N., F. Nedjaoum, D. Guillochon, F. Bureau, P. Arhan, D. Bougie (2002), "Influence of the
extent of hemoglobin hydrolysis on the digestive absorption of heme iron. An in vitro study", Journal
of Agricultural and Food Chemistry, 50 (17): 4969-4973.
64. Mackenzie B., M. D. Garrick (2005), "Iron Imports. II. Iron uptake at the apical membrane in the
intestine", American Journal of Physiology, 289 (6): G981-G986; Hallberg L. (1981), "Bioavailability
of Dietary Iron in Man", Annual Review of Nutrition, vol. 1: 123-147; Sarker S. A., L. Davidsson, H.
Mahmud, T. Walczyk, R. F. Hurrell, N. Gyr, G. J. Fuchs (2004), "Helicobacter pylori infection, iron
absorption, and gastric acid secretion in Bangladeshi children", The American Journal of Clinical Nutri-
tion, 80 (1): 149-153.
65- Monsen E. R. (1988), "Iron nutrition and absorption: dietary factors which impact iron bioavail-
ability", Journal of the American Dietetic Association, 88 (7): 786-90; Pallares I., M. S. Campos, I.
Lopez-Aliaga, M. Barrionuevo, A. E. Gomez-Ayala, M. J. Alferez, S. Hartiti, F. Lisbona (1996), "Sup-
plementation of a cereal-milk formula with haem iron palliates the adverse effects of iron deficiency
on calcium and magnesium metabolism in rats", Annals of Nutrition and Metabolism, 40 (2): 81-90.
66. Hallberg L., L. Rossander-Hulthen, M. Brune, A. Gleerup (1993), "Inhibition of haem-iron ab-
sorption in man by calcium", British Journal of Nutrition, 69 (2): 533-540.
30
Animal by-products (ABPs): origins, uses, and European regulations
interference of the absorption of both heme- and non-heme-Fe have important nutritional
implications.
Also the antioxidant activity of blood protein peptides has been studied and demon-
strated. Xu and others 67 investigated the antioxidant activity of porcine plasma hydrolysate
obtained by pepsin and papain digestion in a peroxidation system of aqueous linoleic acid.
Both the pepsin and papain digested hydrolysates exhibited significant activities against lin-
oleic acid oxidation and good DPPH free radical scavenging ability. Blood has the potential
to serve as a useful source of valuable peptides with antihypertensive, antioxidant, and anti-
bacterial properties. Several bioactive peptides that offer other health benefits such as analge-
sic and antinociception (reduction in pain sensitivity) activities have also been isolated from
blood proteins. Gomes and others 68 provide more detailed information on some of these
peptides that have been isolated from blood. They review the status of the hemoglobin (Hb)
peptide field and highlight recent reports on the identification of a molecular target for a
novel set of Hb peptides, hemopressins, and the implication of these peptides to normal cell
function and disease. The potential therapeutic applications for these Hb-derived hemopres-
sin peptides are also discussed.
Binders or meat glues
Traditionally, binders have been used in meat products to counter the textural and sen-
sorial changes brought about by processing. Binders have a macromolecular structure that
has the capacity to form matrices that retain aroma and nutrients and also entrap large
amounts of water. 69 Devadason and others 70 point out that binders or glues are used to bind
water and fat to stabilize meat emulsions in ground meat products. A particular kind of glue
is represented by a powder substance (meat glue) made from an animal blood-clotting agent
(a transglutaminase enzyme) that connects small pieces of meat. Jacquay et al. 71 have investi-
gated the modifications, as desirability and strength of bind, of meatloaf caused by different
hamburger binders: powdered skim milk, egg binding and powdered soy milk.
67. Xu X., R. Cao, L. He, N. Yang (2009), "Antioxidant activity of hydrolysates derived from porcine
plasma" ', Journal of the Science of Food and Agriculture, 89 (11): 1897— 1903; Alvarez C, M. Rendueles,
M. Diaz (2012), "Production of Porcine Hemoglobin Peptides at Moderate Temperature and Medium
Pressure under a Nitrogen Stream. Functional and Antioxidant Properties", Journal of Agricultural and
Food Chemistry, 60 (22): 5636-5643.
68. Gomes I., C. S. Dale, K. Casten, M. A. Geigner, F. C. Gozzo, E. S. Ferro, A. S. Heimann, L. A.
Devi (2010), ibid. AAPS Journal, 12 (4): 658-669.
69. Chen M. J., C. W. Lin (2002), "Factors affecting the water-holding capacity of fibrinogen/plasma
protein gels optimized by response surface methodology" , Journal of Food Science, 67 (7): 2579-2582.
70. Devadason I. P., A. S. R. Anjaneyulu, Y. Babji (2010), "Effect of different binders on the physico-
chemical, textural, histological, and sensory qualities of retort pouched buffalo meat nuggets", Journal
of Food Science, 75 (1): S31-S35- They studied the functional properties of 4 binders, namely corn
starch, wheat semolina, wheat flour, and tapioca starches, in order to assess their ability to improve the
quality of buffalo meat nuggets processed in retort pouches. Incorporation of corn starch in buffalo
meat nuggets produced more stable emulsion than other binders used. More recently, have investigated
the modifications, as desirability and strenght of bind, of meatloaf caused by different hamburger
binders: powdered skim milk, egg binding and powdered soy milk.
71. Jacquay A., C. Beaver, S. Fisher (201 1), "Alternate hamburger binding methods and their effects on
flavour", Food Chemistry FN 453, Research Project-Fall (www.cfs.purdue.edu/fn/fn453/).
31
Raffaella Leoci
Transglutaminases are a family of proteins that are widely distributed in all living organ-
isms. They are enzymes needed in living animal organisms to repair lesions of body tissues
and create stable structures by extensively cross-linking protein molecules. 72 There has been
a recent surge in findings supporting that dysregulated expression and function of transglu-
taminases contribute to pathological conditions, such as neurodegenerative disorders, tissue
fibrosis, cancer metastasis, celiac disease, and diseases related to defective assembly of the
stratum corneum of the skin.
The TGM1 gene 73 ("transglutaminase- 1"), that provides instructions for making an en-
zyme called transglutaminase- 1, is found in cells that make up the outermost layer of the
skin (the epidermis). The transglutaminase- 1 enzyme is involved in the formation of the
cornified cell envelope, which is a structure that surrounds skin cells and protects against
water loss and infection. The cornified cell envelope is made up of multiple proteins that
are linked to one another (crosslinked). The crosslinking of these proteins is facilitated by
the transglutaminase- 1 enzyme that is essential for the assembly of the cell envelope barrier
in stratified squamous epithelia. It is usually bound to membranes, but to date most studies
with it have involved solution assays. 74
It was also found that the "Transglutaminase-2" (TG2), defined as an enigmatic enzyme
with diverse functions 75 and as a molecular Swiss army knife, 76 is an extremely versatile pro-
tein exhibiting transaminating, protein disulphide isomerase and guanine and adenine nu-
cleotide binding and hydrolyzing activities. TG2 can also act as a protein scaffold or linker.
This unique protein also undergoes extreme conformational changes, exhibits localization
diversity and its multiple biochemical activities account for its involvement in a wide variety
of cellular processes encompassing differentiation, inflammation, cell death, cell migration,
and wound healing.
Transglutaminase-3 (TG3), also known as epidermal Transglutaminase (Tgase E), be-
longs to the family of Transglutaminase enzymes that catalyze the posttranslational modi-
fication of proteins via calcium dependent cross-linking reactions. TG3 is involved in the
formation of the cornified envelope in skin keratinocytes. 77
72. Greenberg C. S., P. J. Birckbichler, R. H. Rice (1991), "Transglutaminases: multifunctional cross-
linking enzymes that stabilize tissues", The FASEB Journal, 5 (15): 3071-7.
73. K polypeptide epidermal type I, protein-glutamine-gamma-glutamyltransferase. See: Kim I. G.,
0. W. McBride, M. Wang, S. Y. Kim, W. W. Idler, P. M. Steinert (1992), "Structure and organiza-
tion of the human transglutaminase 1 gene", The Journal of Biological Chemistry, 267 (11): 7710-7;
Grenard P., M. K. Bates, D. Aeschlimann (2001), "Evolution of transglutaminase genes: identification
of a transglutaminase gene cluster on human chromosome 15ql5- Structure of the gene encoding
transglutaminase X and a novel gene family member, transglutaminase Z", The Journal of Biological
Chemistry, 276 (35): 33066-78.
74. Nemes Z., L. N. Marekov, P. M. Steinert (1999), "Involucrin Cross-linking by Transglutaminase
1. Binding to membranes directs residue specificity", The Journal of Biological Chemistry, 274 (16):
11013-11021.
75- Fesus L., M. Piacentini (2002), "Transglutaminase 2: an enigmatic enzyme with diverse functions",
Trends in Biochemical Sciences, 27 (10): 534-9.
76. Gundemir S., G. Colak, J. Tucholski, G. V. Johnson (2012), "Transglutaminase 2: a molecular
Swiss army knife", Trends in Biochemical Sciences, 1823 (2): 406-19.
77. Zhang J., H. Y. Zhi, F. Ding, A. P. Luo, Z. H. Liu (2005), "Transglutaminase 3 expression in
C57BL/6J mouse embryo epidermis and the correlation with its differentiation", Cell Research, 15 (2):
105-10.
32
Animal by-products (ABPs): origins, uses, and European regulations
The recently introduced synthetic form of these substances develop similar effects in
meat as they have the capacity to form bonds between superficial protein structures of in-
dividual muscle meat pieces. Transglutaminase enzymes act only on proteins by catalyzing
reactions in the formation of covalent bonds between a carboxylamide group of the lateral
chain on a glutamine residue and an amino group of the lateral chain of a Lysine. These
bonds may be formed between proteins of distinct types and origin, as myosins, caseins, soya
globulins, glutens, actins, etc.
As a consequence transglutaminases may be used 78 in various meat processing phases
for bonding a diverse range of food materials such as meat, poultry, dairy, and seafood; from
tumbling and reconstituting cooked hams; to creating protein network structures. Trans-
glutaminase enzymes have an impact on specific meat processing technologies: tumbling
procedures could be shortened, the utilization of phosphates and other binding substances
reduced or completely substituted, and the confection of sausages without membranes,
etc. Even the presence of transglutaminases in raw-fermented sausages, formed by a mix of
coarsely chopped meat and fat particles, can be strengthened by the cohesion of such par-
ticles during ripening. By binding thin cuts of meat together, one can obtain bigger pieces.
Irregular cuts can be "glued" together into a more regular shape and permit even cooking.
In this way it can be useful to glue chicken or bacon or fish skin onto other types of meat
and fish. The enzyme enables food processors to adhere various types of meat together, for
example, as in imitation crabmeat.
British chef Blumenthal 7 '' is credited with the introduction of transglutaminases 80 into
modern cooking. He uses a white powder made by "taking the clotting agents out of pigs"
and cows "blood" in order to clot together chunks of meat. 81
Pares et al 82 have studied the properties of gels obtained from porcine blood plasma
under different pH conditions. Gels from liquid and spray-dried plasma were prepared and
analyzed for water holding capacity (WHC), texture, and microstructure at pH 7.4, 6, 5.5
and 4.5. At low acid pH, such as that of fermented products, plasma is not effective as a gel-
ling agent producing soft and exuding gels. The penetration force of the gel from dehydrated
plasma was always lower than that prepared from liquid plasma where the pH was the same,
but neither the WHC nor the microstructure of gels were affected.
78. Motokia M., K. Segurob (1998), "Transglutaminase and its use for food processing", Trends in
Food Science & Technology, 9 (5): 204-210.
79. Heston Blumenthal owns "The Fat Duck Restaurant", awarded three Michelin stars, restaurant of
the year, and considered best restaurant in the world and best restaurant in the UK.
80. Yokoyama K., N. Nio, Y. Kikuchi (2004), "Properties and applications of microbial transglutamin-
ase", Applied Microbiology and Biotechnology, 64 (4): 447-454.
8 1 . The process begins with sprinkling a teaspoon of powdered transglutaminase on various meat
scraps (sometimes lesser-quality meat) and binding them together with the hands. Roll them up in
plastic wrap. Put in the fridge and 6 hours later, you have an easily-sliced piece of meat that looks
like filet mignon. The meat glue, that has been used in American restaurants for decades, is USDA-
approved and considered safe for the diner. The problem lies with the increased potential for bacteria in
meat that has been created by multiple pieces. The meat glue additive was approved formally in Europe
in 2010, but one year later the arguments against its use are growing.
82. Pares D., E. Saguer, J. Saurina, J. J. Suno, C. Carretero (1998), "Functional Properties of Heat
Induced Gels from Liquid and Spray-Dried Porcine Blood Plasma as Influenced by pH", Journal of
Food Science, 63 (6): 958-961.
33
Raffaella Leoci
Every country in Europe, in recent years, first approved, then banned, and then permit-
ted the use of meat glue or transolutaminases, called also thrombian. In May 19, 2010, the
European Parliament voted to ban the use of a kind of glue 83 as it believes that there is "a
clear risk that meat containing thrombin would find its way into meat products served in
restaurants and other public establishments serving food, given the higher prices that can
be obtained for pieces of meat served as a single meat product". Legislators considered that
consumers should be able to trust that the meat they are buying is a real steak and not simply
pieces of meat glued together. The ban, however, never took effect, as in accordance with
Commission directive 2010/67/EU, 84 and meat glue is permitted for use as a food additive
for reconstituting food.
In the USA, this binder is permitted in products such as blood sausage, blood pudding,
blood soup, and in beef patties. 85 "Beef fibrin" defined as "a component mixture of beef fi-
brinogen and beef thrombin plasma protein used to bind pieces of meat or poultry together"
is permitted up to 10%, provided it is labeled as required. 86
In addition, in the USA, a coating of beef blood is permitted on cured products (e. g.
ham, hamette, etc.), and doubts about its usage, such as those expressed in the EU, have not
been an issue.
Blood flours
Blood flour is dried animal blood, typically cow blood, but it can also be the blood of
any animal that goes through a meat packing plant. Blood is often collected in conjunction
with the slaughtering of animals and subsequently used as a supplemental protein source
for livestock. Normally it is dried and made into blood meal so that it can be handled and
incorporated into feed more easily. The blood is collected after the animals are killed and
then dried to make a powder.
In practice, the withdrawal of blood necessitates a specially designed hollow knife, steri-
lized between each sticking operation, to which a hose is attached. The hose allows the blood
to flow into a container. After bleeding, blood clots in 3-10 minutes. This clotting is caused
by thrombin, which converts soluble fibrinogen in the blood into insoluble fibrin. Antico-
agulants are usually added in the hollow knife. The anticoagulants normally used are either
trisodium citrate (sodium citrate), or citric acid, or a mixture of phosphates, or sodium hex-
ameta phosphate, or sodium acid pyrophosphate, or heparin 87 or oxalates. 88
83. A Nebraska (USA) based company produces a kind of binder that consists of two components:
Fibrinogen i.e. fibrinogen enriched plasma, and Thrombin, an enzyme which initiates the gelling and
the subsequent binding process of meat or fish pieces.
84. Commission Directive 2010/67/EU of 20 October 2010, amending Directive 2008/84/EC laying
down specific purity criteria on food additives other than colours and sweeteners (Official Journal of
the European Union, L. 277/17, 21.10.2010).
85. See: Tarte R. (2009) (Edtr.), Ingredient in Meat Products: Properties, Functionality and Applica-
tions, Springer, New York, pp. 163-4.
86. USDA/FSIS, (2005), Food Standards and Labeling Policy Book. Available: http://www.fsis.usda.
gov/OPPDE/larc/ Policies/Labeling. Policy_Book_082005.pdf
87. Heparin is a natural blood component that helps prevent coagulation in the live animal during
blood circulation. Commercially, it is available in the sodium, lithium, or calcium salt, and it inhibits
the formation of thrombin from prothrombin.
88. Oxalate is poisonous and can not be used for blood which used for food or pharmaceutical uses.
34
Animal by-products (ABPs): origins, uses, and European regulations
Normal bleeding time used in industry is 6' for cattle, 4-5' for sheep, 3-4' for calves,
and 6' for pigs. The quantity of blood collected during bleeding in the slaughter operation
is approximately 50% of the total present in the body since the remaining blood (50%) is
retained in the body's capillary system.
Animal blood is either spray dried as regular whole blood or, after separation, into
plasma and red albumin. Normally blood is dried using one of the following methods:
drum, ring, flash or spray dried. 89 The drying method used has an effect on the digestibility
of the crude protein (CP) of the blood meal, because there is a direct relationship between
the amount of heat applied and the digestibility of the CP. 90 As the amount of heat applied
increases the digestibility of the CP decreases. 91
Blood proteins
Protein malnutrition is a great problem in poor and in developing countries, especially
during the transitional phase of weaning in infants, as it retards their physical and mental
development. Kwashiorkor, 92 more common in very poor countries of some parts of the
world, has been linked to the quality and quantity of protein in maize when it is the sole
source of protein for infants 93 .
Blood protein may be used as a potential source of large quantities of dietary protein,
because it is well balanced in amino acid composition. Since the blood protein is abundant, 94
89. Cook E. M., H. D. DuMont (1971), Process Drying Practice, McGraw-Hill Publishing Co., New
York; Williams-Gardner A. (1991), Industrial Drying, Leonard Hill Books, London.
90. Waibel P. E., M. Cuperlovic, R. E Hurrell, K. J. Carpenter (1977), "Processing damage to lysine
and other amino acids in the manufacture of blood meal" ', Journal of 'Agricultural and Food Chemistry,
25 (1): 171-175.
91. Kats L. J., J. L. Nelssen, M. D. Tokach, R. D. Goodband, T. L. Weeden, S. S. Dritz, J. A. Hansen
(1994), "The effects of spray-dried blood meal on growth performance of the early-weaned pig" ', Jour-
nal of 'Animal Science, 72 (8): 2075-2081.
92. "Kwashiorkor" is a form of malnutrition that occurs when there is not enough protein in the
diet. It often occurs during political unrest, a drought or other natural disaster. These conditions are
responsible for a lack of food, which leads to malnutrition. It is very rare in children in the UE, United
States and in other countries as Japan, South Korea, Australia , etc. where the diet is rich in protein.
93. Of this opinion: Cantor S. M., H. J. Roberts (1967), "Improvement in protein quality in cornbased
foods", Cereal Science Today, 12: 443-445, 460-462; Banigo E. O. I., A. I. Adeyemi (197 '5), A comparative
study of the commercial practice of traditional ogi manufacture using high-lysine ( opaque-2) corn and normal
corn, Proceedings of the 10th International Congress of Nutrition, Kyoto, Japan, p. 402; Akinrele I. A.,
C. C. A. Edward (1971), "An assessment of the nutritive value of maize soya mixture. «Soya-ogi» as a
weaning food in Nigeria", British Journal of Nutrition, 26 (2): 177-185; Makinde M. A., P. A. Lachange
(1989), "Optimization of protein nutritive value of ogi", Nigerian Journal of Nutrition Science, 10: 85-93;
Mottern H. H., T. S. De Buckle, C. Pardo (1970), "Protein enrichment of Colombian corn cakes", Cereal
Science Today, 15: 108-112; FAO, Maize in Human Nutrition, Food and Agriculture Organization of the
United Nations, FAO Food and Nutrition Series, n. 25,1992, pp. 81-131.
94. It has been estimated that only in China about 1,5 millions of tons of porcine blood, produced
annually until 2006, has a proteine content equivalent to that 2 millions of tons of meat (See: Wang J.
Z., M. Zhang, F. Z. Ren, B. Z. Han, L. Wang, S. W. Chen, A. Humera (2007), "Changes of chemical
and nutrient composition of porcine blood during fermentation by Aspergillus oryzae", World Journal
of Microbiology and Biotechnology, 23: 1393-1399). In Italy we have estimated that the annual produc-
tion of bovine blood is about 40,000 t and of porcine blood is roughly 150,000 t.
35
Raffaella Leoci
cheap, readily available, and has a proven track record in animal nutrition, it has been sug-
gested for use as protein supplements in infant formula to tackle protein malnutrition.
Five experiments were conducted by Pierce and others, 95 to evaluate the effects of di-
etary spray-dried porcine plasma (SDPP) and spray-dried bovine plasma (SDBP) and their
various molecular weight fractions on performance of pigs weaned at approximately 14-21
days of age. These studies verified that the immunoglobulins, primarily immunoglobulin G,
are the major components in plasma that stimulates growth in early-weaned pigs. Similar
results have been observed in newly weaned mice, 96 indicating that the benefits of including
blood proteins in the diet are not species-specific. Thomson and others conclude that mice
respond to dietary inclusion of spray-dried porcine plasma protein (SDPP) with increases in
average daily feed intake (ADFI), Average daily gain (ADG), and gain-to-feed ratio (G/F)
during the period immediately after weaning may, therefore, serve as appropriate models for
pig responses to SDPP.
The adequacy of blood proteins as protein supplements has also been demonstrated
by Oshodi and others, 97 who investigated in vitro multienzyme protein digestibility of an
infant-weaning food produced from maize flour and bovine blood proteins using the pH-
stat and pH-drop procedures. Their results indicated an improved digestibility when blood
protein concentrate was added to the maize. After blending with the blood protein con-
centrate, the level of available iron was double that of maize flour alone. The bovine blood
powder concentrate used was so bland in taste that its addition to the maize flour did not
alter the overall flavor.
Recent physiological and biochemical research has shown that the protein in food
not only furnishes amino acids but also provides bioactive peptides after digestion or food
processing. Clare and Swaisgood 98 have reviewed the scientific literature and attempted to
stimulate consideration of the continued use of bioactive peptides and their expanded devel-
opment as a commercial product. Several applications have already evolved. Potentially, the
addition of bioactive peptides to food products could improve consumer safety as a result
of their antimicrobial properties. Bioactive peptides may function as health care products,
providing therapeutic value for either treatment of infection or prevention of disease. As a
consequence, bioactive peptides produced from both plant and animal sources are being
investigated and have been reported to have antinematodal, 99 antiviral and antibacterial, 100
95. Pierce J. L„ G. L. Cromwell, M. D. Lindemann, L. E. Russell, E. M. Weaver (2005), "Effects of
spray-dried animal plasma and immunoglobulins on performance of early weaned pigs", Journal of
Animal Science, 83 (12): 2876-2885.
96. Thomson J. E., E. E. Jones, E. J. Eisen (1994), "Effect of spray-dried porcine plasma protein on
feed intake, growth rate, and efficiency of gain in mice", Journal of "Animal Science, 72 (10): 2690-2695.
97. Oshodi A. A., R. M. Beames, S. Nakai (1997), "In vitro protein digestibility, amino acid profile
and available iron of infant weaning food prepared from maize flour and bovine blood", Food Research
International, 30 (3/4): 193-197.
98. Clare D. A., H. E. Swaisgood (2000), "Bioactive milk peptides: a prospectus", Journal of Dairy
Science, 83 (6): 1187-1195.
99. The antinematodal activity and mechanism of a 23-mer antimicrobial peptide, PMAP-23, derived
from pig myeloid was investigated by Yoonkyung Park, Seung-Hwan Jang, Dong Gun Lee, Kyung-Soo
Hahm (2004), (in "Antinematodal effect of antimicrobial peptide, PMAP-23, isolated from porcine
myeloid against Caenorhabditis elegans' ', Journal of "Peptide Science, 10 (5): 304-31 1).
100. Jenssen H„ P. Hamill, R. E. W. Hancock (2006), "Peptide Antimicrobial Agents", Clinical Micro-
36
Animal by-products (ABPs): origins, uses, and European regulations
opioid, 101 antioxidant, 102 antitumor, 103 and angiotensin I-converting enzyme (ACE) inhibi-
tory (anti-hypertensive) 104 activities.
The studies previously listed support the use of bovine blood proteins as efficient protein
supplements in cereal based weaning and infant diets as a measure to tackle protein malnu-
trition problems.
The suspicion that blood provides a haven for pathogens and toxic metabolites is a
strong reason why some people avoid consuming blood. It is however necessary to remem-
ber, that in general, foods of animal origin are easily contaminated with microorganisms,
often pathogens, due to poor handling and processing. 105
biology Review, 19 (3): 491-51 1; Lee D. G., D.-H. Kim, Y. Park, H. K. Kim, H. N. Kim, Y. K. Shin, C.
H. Choi, K. -S. Hahm (2001), "Fungicidal effect of antimicrobial peptide, PMAP-23, isolated from
porcine myeloid against Candida albicans", Biochemical and Biophysical Research Communications, 282:
570-574; Yu P. L., S. D. Choudhury,, K. Afiren (2001), "Purification and characterization of the anti-
microbial peptide, ostricacin", Biotechnology Letters, 23 (3): 207—210.
101. Perpetuo E. A., L. Juliano, I. Lebrun (2003), "Biochemical and pharmacological aspects of two
bradykinin-potentiating peptides obtained from tryptic hydrolysis of casein" , Journal of Protein Chem-
istry, 22 (7/8): 601-606.
102. Tri Agus Siswoyo, Eka Mardiana, Kyun Oh Lee, Keizo Hoshokawa (201 1), "Isolation and Char-
acterization of Antioxidant Protein Fractions from Melinjo (Gnetum gnemon) Seeds" ', Journal of Ag-
ricultural and Food Chemistry, 59 (10): 5648-5656; Se-Kwon Kim, Yong-Tae Kim, Hee-Guk Byun,
Kyung-Soo Nam, Dong-Sik Joo, F. Shahidi (2001), "Isolation and characterization of antioxidative
peptides from gelatine hydrolysate of Alaska pollack skin" , Journal of Agricultural and Food Chemistry,
49 (4): 1984-199; Je-Ruei Liu, Ming-Ju Chen, Chin-Wen Lin (2005), "Antimutagenic and antioxi-
dant properties of milk-kefir and soymilk-kefir" , Journal of Agricultural and Food Chemistry, 53 (7):
2467-2474; Pratt D. E., C. Di Pietro, W. L. Porter, J. W. Giffee (1982), "Phenolic antioxidants of soy
protein hydrolyzates" , Journal of Food Science, 47 (1): 24-35; Esaki H., R. Watanabe, H. Onozaki, S.
Kawakishi, T. Osawa (1999), "Formation mechanism for potent antioxidative o-dihydroxyisoflavones
in soybeans fermented with Aspergillus saitoi", Bioscience, Biotechnology, and Biochemistry, 63 (5): 851-
858; Chen H. M., K. Muramoto, F. Yamauchi, K. Nokihara (1996), "Antioxidant activity of designed
peptides based on the antioxidative peptide isolated from digests of a soybean protein", Journal of
Agricultural and Food Chemistry, 44 (9): 2619-2623; Tong L. M„ S. Sasaki, Julian D. McClements,
E. A. Decker (2000), "Mechanisms of the antioxidant activity of a high molecular weight fraction of
whey", Journal of Agricultural and Food Chemistry, 48 (5): 1473-1478.
103. Zeiger E. (1987), "Carcinogenicity of mutagens: Predictive capability of the Salmonella muta-
genesis assay for rodent carcinogencity", Cancer Research, 47 (5): 1287-1296; Qureshi A., P. L. Colinb
D. J. Faulkner (2000), "Microsclerodermins F-I, antitumor and antifungal cyclic peptides from the
lithistid sponge Microscleroderma sp.", Tetrahedron, 56: 3679-3685; Shiomi M., K. Sakaki, M. Mu-
rofushi, K. Aibara (1982), "Antitumor activity in mice of orally administered polysaccharide from kefir
grain", Japanese Journal of Medical Science dr Biology, 35: 75-80.
104. Matsui T., Akiko Yukiyoshia, Shima Doib, Hiroyuki Sugimotob, Hideo Yamadab, Kiyoshi Mat-
sumotoa (2002), "Gastrointestinal enzyme production of bioactive peptides from royal jelly protein
and their antihypertensive ability in SHR", The Journal of Nutritional Biochemistry, 13 (2): 80-86;
Arihara K, Y. Nakashima, T. Mukai, S. Ishikawa, M. Itoh (2001), "Peptide inhibitors for angiotensin
I-converting enzyme from enzymatic hydrolysates of porcine skeletal muscle proteins", Meat Science,
57 (3): 319-324; Kapela R., A. Chabeaua, J. Lesagec, G. Rivierec, R. Ravallec-Plea, D. Lecouturiera,
M. Wartelled, D. Guillochona, P. Dhulster (2006), "Production, in continuous enzymatic membrane
reactor, of an anti-hypertensive hydrolysate from an industrial alfalfa white protein concentrate exhib-
iting ACE inhibitory and opioid activities", Food Chemistry, 98 (1): 120-126.
105. Al-Bachir M„ A. Mehio (2001), "Irradiated luncheon meat: microbiological, chemical and sen-
37
Raffaella Leoci
Manufacturers and processors have instituted measures to guarantee the safety of these
blood proteins. In the UE, article 10 of Regulation (EC) No 1069/2009 of 21 October
2009 and in the US, Federal Regulation 9 CFR 310.20 monitors blood that enters the food
chain, ensuring that it originates from official establishments whose livestock and carcasses
have passed inspection.
Blood sausages
The traditional use of blood, however, is generally restricted to such products as blood
sausages where the black color is both expected and acceptable. When plasma is used in
meat sausages, because of its gelling properties, it will decrease shrinkage and increase yield,
and the texture of the finished product becomes more rigid. Meat sausage, produced with
emulsification of fat by plasma, is able to retain the fat during its heating.
Clarifiers
A clarifying or fining agent makes wine clear by removing proteins from the wine. The
agents eventually settle out of the wine. Different proteins serve as clarifying agents depend-
ing upon both the type of wine and the desired flavor. Some clarifiers are animal-based
products, while others are earth-based. Wine may be refined either with eggs, milk, or dried
blood powder. Although blood of large mammals may serve as a clarifier in some old Medi-
terranean countries, its use is forbidden in wine from either the United States or Europe.
Curing agents
Curing is the treatment of muscle meat with common salt or sodium chloride (NaCl)
and sodium nitrite (NaN0 2 ). In the past, curing was mainly applied to extend the storage
life of entire pieces of muscle meat by using the preserving effects of common salt and to
a lesser extent sodium nitrite. In modern meat processing, the storage life is less important
since more efficient meat preservation methods, in particular cooling and freezing, are avail-
able, and curing is now applied to achieve a pink-red color as well as a typical flavor and taste
in processed meat products.
Although sodium nitrite continues to be used as a common food additive, its use has
come under criticism in recent years because it has a tendency to react with amines, amides,
amino acids and related compounds present in the meat to produce carcinogenic nitroso-
amine compounds. As a consequence, efforts are being made to reduce the amount of so-
dium nitrite used in the curing system and to develop alternative methods of curing that
avoid the use of sodium nitrite altogether. 106
Nowadays, no single capable agent has been identified to replace nitrite, as it is extreme-
ly difficult to find a single compound that can fully reproduce the multifunctional role of ni-
trite (as a colorant, antioxidant, and antimicrobial). 107 One of the colorants suggested for use
sory characteristics during storage", Food Chemistry, 75 (2): 169-175.
106. PeggR. B., F. Shahidi, Nitrite Curing of 'Meat: The N-Nitrosamine Problem and Nitrite Alternatives,
Food & Nutrition, Inc., Trumbull, Connecticut, USA, pp. 209-254.
107. Nollet L. M. L., F. Toldra (Edts) (2006), Advanced Technologies For Meat Processing, CRC Press -
Taylor & Francis Group, NW, pp. 309-328.
38
Animal by-products (ABPs): origins, uses, and European regulations
in a composite sodium nitrite-free cocktail for meat curing is Cooked Cured Meat Pigment
(CCMP), prepared by reacting bovine or porcine red blood cells with a nitrosating agent. 108
According to Shahidi and Pegg, 109 the application of this CCMP in different formulations
to comminuted and solid cuts of muscle foods has showed that color, oxidative stability and
flavor of the treated samples were similar to those of their nitrite-cured counterparts. Shahidi
et al. 110 have also investigated the effects of 5 and 10 kGy irradiation on the color and oxida-
tive stability of meats treated with nitrite or a nitrite-free curing system.
Dyes and color enhancers
Color, as it is well known, is one of the main factors used by consumers when evaluating
the quality and freshness of food products, especially meat products. The growing aversion
to some artificial food dyes has resulted in the substitution of artificial colorants with natural
ones. Stabilized hemoglobin (frozen or powder hemoglobin), 111 obtained from porcine or
bovine blood, represents a good source of natural red colorant and is also an iron supple-
ment for meat products in some countries. In order to stabilize hemoglobin or to prevent
hemoglobin auto-oxidation during drying and subsequent storage, Salvador et al. 112 suggest
the use of chelating agents, such as nicotinic acid (NA, 2% w/v) or nicotinamide (Nam,
2.5% w/v) along with glucose, as reducing agents (G, 10% w/v) which can be combined
with fresh porcine hemoglobin in order to stabilize its red color during spray-drying and
during powder storage at room temperature. These chelating agents have the ability to form
complexes with the heme moiety. From the results, it can also be concluded that glucose is
the main contributor to the color stabilization of the hemoglobin powder.
Mancini and Hunt" 3 recently updated research regarding numerous ante-mortem and
post-mortem factors that influence meat color. They noted that carbon monoxide saturation
of hemoglobin resulted in conversion into the more stable carboxyhemoglobin and has been
proposed as an alternative method to stabilize hemoglobin.
108. Shahidi E, R. B. Pegg (i990), "Colour characteristics of cooked cured-meat pigment and its ap-
plication to meat", Food Chemistry, 38 (i): 6i-68.
109. Shahidi E, R. B. Pegg (1992), "Nitrite-free meat curing systems: Update and review", Food Chem-
istry, 43 (3): 185-191.
110. Shahidi E, R. B. Pegg, K. Shamsuzzaman (1991), "Color and Oxidative Stability of Nitrite-Free
Cured Meat after Gamma Irradiation' ', Journal of Food Science, 56 (5): 1450-1452.
111. Spray-drying haemoglobin is a good way to preserve the red blood cell fraction, better than freeze
drying (See: Saguer E., S. Altarriba, C. Lorca, D. Pares, M. Toldra', C. Carretero (2003), "Colour
stabilization of spray-dried porcine red blood cells using nicotinic acid and nicotinamide", Food Science
and Technology International, 9 (4): 301-307).
1 12. Salvador E, M. Toldra, D. Pares, C. Carretero, E. Saguer (2009), "Color stabilization of porcine
hemoglobin during spray-drying and powder storage by combining chelating and reducing agents",
Meat Science, 83 (2): 328-333.
113. Mancini R. A., M. C. Hunt (2005), "Current research in meat color", Meat Science, 71 (1):
100-121. See also: Knock R. C, M. Seyfert, M. C. Hunt, M. E. Dikeman, R. A. Mancini, J. A.
Unruh, J. J. Higgins, R. A. Monderen (2006), "Effects of potassium lactate, sodium chloride, sodium
tripolyphosphate, and sodium acetate on colour, colour stability, and oxidative properties of injection-
enhanced beef rib steaks", Meat Science, 74 (2): 312-318.
39
Raffaella Leoci
Egg albumin substitute
Blood albumin is used as a substitute for egg albumin in food. Albumin is a type of
globular protein that has a roughly spherical structure and is water-soluble. Many types
can be found in the natural world, and two of the most familiar examples can be found
in egg whites (known as ovalbumin) and in human blood. Albumin is vitally important
to the health and well being of many organisms. It is an important component of life be-
cause it transports essential fatty acids from adipose tissue to muscle tissue. The protein also
contributes to the regulation of osmosis, helping to transport hormones, drugs, and other
substances through the blood.
When heated, albumin and other proteins tend to coagulate; they transform, recom-
bining in a new configuration and turn white and opaque. This property mystified ancient
alchemists, because, generally, substances become liquid when heated. The protein in egg
white albumin helps baked goods hold their structure, and the same egg white albumin is
also used for purification, as it tends to trap and store impurities. In addition, albumin is
used to treat people with certain types of poisoning, since it binds to the toxin.
In the preparation of bakery products, in practice, many additives are used to improve
the properties of the dough: eggs, powdered soy, milk, etc. Jacquay and coll. 114 have investi-
gated the modifications, such as desirability and strength, in meatloaf as caused by different
hamburger binders: egg, powder skim milk and powdered soymilk. Bovine blood plasma is
a useful source of high-quality protein and exhibits many useful properties. Blood plasma,
used in bakery products instead of egg albumin, has good foaming and leavening properties.
Adding plasma (no more of 2%) to bread flour encourages a considerable higher loaf volume
and also increases the protein quality. Thus the bread contains about 1 5% more protein and
around 75% more lysine. Spray-dried plasma can be used as an egg substitute. Plasma levels
greater than 2% darkened the crust and make the texture of bread more open and coarse.
Many researches 115 found that in cake making about 30% of whole egg could be replaced
with blood plasma. In those studies, full replacement with blood plasma did not quite pro-
duce the same volume, crumb structure and profile as egg white.
Raeker et al 116 have investigated the cake-baking properties of egg white and blood
plasma. They concluded that egg white produced slightly larger cake volume, a significantly
more crowned profile, and a finer crumb structure than did blood plasma. Among the blood
plasma proteins, fibrinogen produced the smallest cake volume. Albumin, the protein in
blood plasma, had cake-baking properties inferior to those of whole blood plasma.
114. Jacquay A., C. Beaver, S. Fisher (2011), ibid., Research Project-Fall (www.cfs.purdue.edu/fn/
fn453/).
115. Brooks J., P. W. Ratcliff (1959), "Dried bovine plasma. I. Storage of spray-dried plasma and the
freeze-concentration of liquid plasma", Journal of the Science of Food and Agriculture, 10 (9): 486-495;
Khan M. N., L. W. Rooney, C. W. Dill (1979), "Baking properties of plasma protein isolate", Journal
of Food Science, 44 (1): 274-76; Johnson L. A., E. F. Havel, R. C. Hoseney (1979), "Bovine plasma as
a replacement for egg in cakes", Cereal Chemistry, 56: 339-41; Lee C. C, L. A. Johnson, J. A. Love,
S. Johnson (1991), "Effects of processing and usage level on performance of bovine plasma as an egg
white substitute in cakes", Cereal Chemistry, 68 (1): 100-104.
116. Raeker M. O., L. A. Johnson (1995), "Cake-Baking (High-Ratio white Layer) Properties of Egg
white, Bovine Blood Plasma, and their Protein Fractions", Cereal Chemistry, 72 (3): 299-303.
40
Animal by-products (ABPs): origins, uses, and European regulations
Emulsifier
An emulsifier or emulsifier agent is a substance that will hold another substance sus-
pended so that the solution is stable. Emulsifiers are utilized in emulsified meat to bind meat
proteins, fat and water in a stable emulsion. In the food industry, among the emulsifiers,
casein and its salt derivatives are widely used. But, because of its high processing costs, it
is expensive. Proteins are also common emulsifying agents as they occur naturally and are
generally widely available, non-toxic and inexpensive. 117 According to some research, protein
isolate produced from food industry by-products may have potential as an emulsifying agent
in the food industry. Blood proteins have been found to have emulsifying properties that
are comparable, and often, superior to those of casein and can therefore replace casein as an
emulsifying agent. Silva et al. used two methods for extraction of globin from bovine blood,
in order to test it in terms of efficiency, protein recovery and solvent residues. 118
The evaluation of the capacity of blood proteins in either forming or stabilizing emul-
sions is highly important from an industrial point of view since the manufacture of several
foods, such as mayonnaise, pates and sausages involves an emulsification process. Moreover,
the incorporation of proteins in food products may increase their nutritional value. On the
other hand, considering that the action of proteins as emulsifiers is complex and depends
on different factors (protein concentration, oil type, velocity and length of mixture, among
others), it is important to study the behavior of proteins under different conditions. 1 " Biz-
zotto et al. 120 have evaluated the emulsifying properties of a blood protein. They studied the
effect of the pH and of the tryptic hydrolysis on the emulsifying properties of bovine globin
and determined the emulsifying capacity (EC), the emulsifying activity index (EAI), and the
emulsion stability (ES) at pH varying from 3.0 to 8.0 and employing hydrolysis times from
5 to 60 minutes. They obtained the highest values for EC and ES at pH 5.0 and 6.0, respec-
tively, corresponding to the range of high protein solubility. The best results for the emulsify-
ing properties of bovine globin, extracted by the acidified acetone method, was observed in
the acid region (pH from 3.0 to 6.0), where this protein is highly soluble.
Finally, blood proteins (hemoglobin), used as emulsifiers, have the further advantage of
providing a good source of heme iron. 121 The success of blood-fortified foods in addressing iron
117. Matsumura Y., K. Matsumiya (2012), Chapter 5, Proteins-Peptides as Emulsifying Agents, in S.
Navam, Hettiarachchy, K. Sato, M. R. Marshall, A. Kannan, Food Proteins and Peptides Chemistry,
Functionality, Interactions, and Commercialization, CRC Press, Florida (USA), pp. 125-150.
118. Silva J. G., H. A. Morais, M. P. C. Silvestre (2003), "Comparative study of the functional proper-
ties of bovine globin isolates and sodium caseinate", Food Research International, 36 (1): 73-80.
119. Tybor P. T., C. W. Dill, W. A. Landmann (1973), "Effect of descolorization and lactose incorpora-
tion on the emulsification capacity of spray-dried blood protein concentrates", journal of Food Science,
38 (6): 4-6; Crenwelge D. D„ C.W. Dill, P.T. Tybor, W. A. Landmann (1974), "A comparison of the
emulsification capacities of some protein concentrates", Journal of Food Science, 39 (910): 175-177;
Gauthier S. E, P. Paquin, Y. Pouliot, S. Turgeon (1993), "Surface activity and related functional prop-
erties of peptides obtained from whey proteins' ', Journal of 'Dairy Science, 76 (1): 321-328.
120. Schaper Bizzotto C, M. Capobiango, M. Pinto Coelho Silvestre (2005), "Evaluation of Func-
tional Properties of a Blood Protein", Pakistan Journal of Nutrition, 4 (1): 11-16.
121. As bovine blood has the largest amount of heme iron than any animal source (see: Kikafunda
J. K„ P. Sserumaga (2005), ibid., African journal of food agriculture nutrition and development, 5 (1):
1-18) and is readily available, it has therefore been suggested for the fortification of staple foods.
41
Raffaella Leoci
deficiency has been reported by several researchers. 122 Compared to egg and casein, that are
potent allergens that are among the "big eight" allergens 123 covered under the "Food Allergen
Labeling and Consumer Protection Act" (FALCPA) law 124 , blood proteins present no problem.
Fat replacers
Fat, besides its nutritional function, plays an important role in the diet as a source of es-
sential fatty acids and energy; as an enhancer of tenderness, palatability and juiciness; and as
a binder of processed meats. As empirical data 125 has shown a correlation between dietary fat
and cardio-vascular disease and some types of cancers, consumers have reacted by reducing
dietary fat in their food. 126 In response to this trend, the food industry has developed an as-
sortment of low-fat meat products. But given the sensory role played by fats, replacing their
presence in food products is a difficult task. Many researchers 127 have shown blood proteins
122. According to Nissenson A. R., C. Charytan (2003) (in "Management of Comorbidities in Kidney
Disease in The 21St Century: Anemia and Bone Disease", Kidney International, 64: S64-S71), "al-
though oral non-heme iron is infrequently sufficient to maintain iron stores in hemodialysis patients,
recent studies suggest that heme-iron may be more useful in this regard. Heme-iron is absorbed to
a greater extent than non-heme iron, and is better tolerated. Small studies have shown that when
heme-iron is administered, less parenteral iron and lower doses of erythropoietin (EPO) are needed
to maintain target haemoglobin"; See also: Hurrell R. F. (1997), "Preventing iron deficiency through
food fortification", Nutrition Reviews, 55 (6): 210-22; Nissenson A. R., J. S. Berns, P. Sakiewicz, S.
Ghaddar, G. M. Moore, R. B. Schleicher, P. A. Seligman (2003), "Clinical evaluation of heme iron
polypeptide: sustaining a response to rHuEPO in hemodialysis patients", American Journal of Kidney
Diseases, 42 (2): 325-330; Seligman P. A, G. M. Moore, R. B. Schleicher (2000), "Clinical studies of
hip: An oral heme-iron product", Nutrition Research, 20 (9): 1279-1286
123. A group of the eight major allergenic foods is often referred to as the Big-81 and comprises milk,
eggs, fish, crustacean shellfish, tree nuts, peanuts, wheat and soybean. These foods account for about
90% of all food allergies in the USA.
124. The "Food Allergen Labeling and Consumer Protection Act" of 2004 (FALCPA) (Public Law
108-282) was enacted in August 2004, and addresses, among other issues, the labeling of foods that
contain certain food allergens.
125. See among others: Takashi Sugimura (1997), "Food and cancer prevention", Cancer Letters, 114
(1-2): 3-5; Greenwalda P., C. K. Cliffordb, J. A. Milner (2001), "Diet and cancer prevention", Euro-
pean Journal of Cancer, 37 (8): 948-965; DAvanzo B., E. Negri, A. Gramenzi, S. Franceschi, F. Paraz-
zini, P. Boyle, C. La Vecchia (1991), "Fats in seasoning and breast cancer risk: an Italian case-control
study", European journal of Cancer and Clinical Oncology, 27 (4): 420-423; Ferguson L. R. (2010),
"Meat and cancer", Meat Science, vol. 84 (2): 308-313; Hooper L., C. D. Summerbell, J. P. T. Higgins,
R. L. Thompson, N. E. Capps, G. Davey Smith, R. A. Riemersma, S. Ebrahim (2001), "Dietary fat
intake and prevention of cardiovascular disease: systematic review", BMJ, 322 (7289): 757-763.
126. Colmenero F. J. (1996), "Technologies for developing low-fat meat products", Trends in Food
Science & Technology, 7 (2): 41-48.
127. Read more: Totosaus A., R. H. Alfaro-Rodriguez, M. L. Perez-Chabela (2004), "Fat and sodium
chloride reduction in sausages using K-carrageenan and other salts", International Journal of Food Sci-
ences and Nutrition, 55 (5): 371-380; Tokusoglu O., M. K. Unal (2003), "Fat Replacers in Meat Prod-
ucts", Pakistan Journal of Nutrition, 2: 196-203; Joly G., B. Anderstein (2009), Starches, Ingredients in
Meat Products, Springer- Verlag, New York, pp 25-55; Viana F. R., V. D. M. Silva, F. M. Delvivo, C. S.
Bizzotto, M. P. C. Silvestre (2005), "Quality of ham pate containing bovine globin and plasma as fat
replacers", Meat Science, 70 (1): 153-160; Viana F. R., C. S. Bizzotto, D. R. Dias, A. L. Oliveira, M. P.
C. Silvestre (2004), "Bovine Blood Constituents as Fat Replacers in Ham Pate", Food Technology and
42
Animal by-products (ABPs): origins, uses, and European regulations
can replaced fat in meat products, while at the same time having the advantage of reducing
costs and the caloric content of food.
Imitation seafood
Surimi (from the Japanese 128 "ground meat") is a fish-based food product that has been
pulverized to a thick paste. When cooked it has the appearance of a rubbery food item. In
general, surimi is sold as imitation crab or mock crab in USA and as seafood sticks, crab
sticks, or fish sticks in Europe. Imitation crabmeat is a seafood product made by blending
processed fish with various texturizing ingredients, flavorants, and colorants resulting in a
gel. 129 This gel can be shaped and cut into thin strips which, when rolled together, mimic the
texture of real crabmeat.
Depending on the desired texture and flavor of the surimi product, the gelatinous paste
is mixed with differing proportions of additives such as egg white, starch, salt, vegetable
oil, humectants, sorbitol, sugar, soy protein, seasonings, and enhancers such as transglu-
taminases and monosodium glutamate. The coloring for imitation crabmeat is made using
compounds like carmine, caramel, paprika, and annato extract.
Stabilizers
Stabilizers are substances or chemicals that allow food ingredients, which do not mix
well, to remain in a homogenous state after blending. Transglutaminase is widely used in sea-
food, surimi products, meat products, noodles/pasta, dairy products, baked goods, whipped
toppings 13 " and so on. It has great potential to improve the firmness, elasticity, viscosity, heat
stability, and water-holding capacity of prepared foods through mild enzyme reaction. 131
b) Feed (blood meal)
Blood can be collected during the slaughter of various livestock species (cattle, pigs,
chickens, etc.) and is usually dried and made into blood meal so that it can be handled and
incorporated into animal rations and can be used as a stabilizer for fat in bone meal. The
yield of blood meal 132 from whole blood is approximately 20%.
Biotechnology, 42 (1): 5-10; Carballo J., G. Barreto, F. J. Colmenero (1995), "Starch and Egg White
Influence on Properties of Bologna Sausage as Related to Fat Content" ', Journal of Food Science, 60 (4):
673-677; Abiola S. S., S. W. Adegbaju (2001), "Effect of substituting pork backfat with rind on qual-
ity characteristics of pork sausage", Meat Science, 58 (4): 409-12.
128. The Japanese have been using surimi-based products for about a thousand years. Traditionally
called "kamaboko", the first recorded surimi manufacturing procedure was found in a Japanese cook-
book written in 1528.
129. Lean meat from fish (or land animals) is first separated or minced. Then it is rinsed many times
to eliminate undesirable odours and pulverized to form a gelatinous paste.
130. Whipped toppings are concoctions of sugar, stabilizers, and some substance approximating cream.
131. As a review see: Kuraishia C, K. Yamazakia, Y. Susa (2001), "Transglutaminase: its Utilization in
the Food Industry", Food Reviews International, 17 (2): 221-246.
132. Blood run through a decanter to separate the coagulated blood into pre-dewatered blood meal
and blood water, which is released during coagulation. The blood meal is then cooked with stirring to
avoid clumping. Sometimes it is added lime (70% calcium oxide), at the 0.5-1.5% level, to increase
storage life and to decrease the odour released during drying. Blood mixed with lime has a rubbery
43
Raffaella Leoci
Blood meal contains mostly protein and is used as a source of protein to supplement
diets based on forages, plant by-products and cereal grains. It has been shown to be a satis-
factory replacement for other supplemental protein sources in various animal productions
including dairy cattle, beef cattle, sheep, pigs, poultry, various fish species, and silkworms. It
is also a good source of most of the trace minerals. Blood meal is rich source of lysine but is
deficient in tryptophan and isoleucine.
Blood meal is often hydroscopic and needs to be dried to less than 10-12% moisture
and stored in a dry place. There are different ways to prepare blood meal: the drying method
used (solar, oven, drum, flash, spray drying, etc.) has different effects on the digestibility of
the blood meal proteins. There is an inverse relationship between the amount of heat applied
and protein digestibility: lysine content decreases when the amount of heat increases. 133
It is worth noting, however, that the advent of "mad cow" disease (Bovine Spongiform
Encephalopathy or BSE) raised international concern about the safety of feeding rendered
cattle to cattle. Since the discovery of mad cow disease in the USA, EN and other countries,
governments have taken action to restrict certain parts of cattle that can be fed back to cattle.
The rise in animal factories over the last 50 years has led to a system that is out of control.
However, most animals are still allowed to eat animal by-products from their own spe-
cies. Pig carcasses can be rendered and fed back to pigs, cattle can be fed cow blood and some
other cow parts, chicken carcasses can be rendered and fed back to chickens, and so on. Also,
cosmetics and dietary supplements cannot be manufactured using potentially infectious cow
parts. In recent years, some supplements have claimed to include cow brains.
c) Fertilizers
Blood is useful for modifying the pH of the soil and to add nitrogen. It is also useful to
improve the soil texture. Its disadvantage is the possibility of attracting rats and vermin when
consistency and can be stored at 20° C for 24 hours without spoilage (it contains 80-85% moisture). In
order to remove moisture, the dark-brown cooked product is pressed and dried to the desired moisture
level. The dried product is then ground and used as feed (80% protein). Spray-dried blood can also
be used as an adhesive, in asphalt emulsions, in insecticides, in ceramics, and as a substitute for egg
albumin when colour is not important. The dried product is often heated to 100° C for 30 minutes,
cooled, and stored in airtight containers.
133. See: Johns D. C, C. K. Low, J. R. Sedcole, M. P. Gurnsey, K. A. C. James (1987), "Comparison
of several in vivo digestibility procedures to determine lysine digestibility in poultry diets containing
heat treated meat and bone meals", British Poultry Science, 28 (3): 397—406; Wang X., C. M. Parsons
(1998), "Bioavailability of the Digestible Lysine and Total Sulfur Amino Acids in Meat and Bone
Meals Varying in Protein Quality", Poultry Science, 77 (0): 1003-1009; Batterham E. S., R. E. Darnell,
L. S. Herbert, E. J. Major (1986), "Effect of pressure and temperature on the availability of lysine in
meat and bone meals as determined by slope-ratio assays with growing pigs, rats and chicks and by
chemical techniques", British Journal of Nutrition, 55 (2): 441-453; Batterham E. S., R. F. Lowe, R.
E. Darnell (1986), "Availability of lysine in meat meal, meat and bone meal and blood meal as deter-
mined by the slope-ratio assay with growing pigs, rats and chicks and by chemical techniques", British
Journal of Nutrition, 55 (2): 427-440; Lewis A. J., D. H. Baker (1995), Bioavailability of D-Amino acids
and DL-Hydroxy Methionine, in C. B. Ammerman, D. H. Baker, and A. J. Lewis (Edtrs), Bioavailabil-
ity of Nutrients for Animals: Amino Acids, Minerals, and Vitamins, Academic Press, London (UK)-San
Diego (USA), pp. 67-81; Parsons C. M., E Castanon, Y. Han (1997), "Protein and amino acid quality
of meat and bone meal", Poultry Science, 76 (2): 361-368.
44
Animal by-products (ABPs): origins, uses, and European regulations
spread on the soil. The dried product is used as fertilizer (12% nitrogen, 0.22% phosphorus,
and trace elements). It is usually mixed with super phosphates to make a compounded fertilizer.
Gagnon and Berrouard 134 evaluated the potential of different organic wastes from the agri-
food industry for growing greenhouse tomato transplants. The organic materials were thor-
oughly mixed with a peat-compost growing medium prior to transplanting. Meal from blood,
feathers, meat, crab shells, fish, cottonseed and whey by-products produced the best growth,
significantly increasing the shoot dry weight by 57-83% compared with non-fertilized plants.
Ciavattaa et al. 135 had incubated blood meal in the soil for one year, following the evolu-
tion of the organic matter. The results showed that only a part (about 75%) of the organic
carbon (C) and of the organic N mineralized and that the remaining C was transformed into
humified compounds. The availability of Fe increased during the incubation, probably due
to the progressive degradation of the prosthetic group and the successive chelation of the Fe
from the humic substances.
In a laboratory incubation Hartz and Johnstone 136 compared the rate of net nitrogen
mineralization (N min ) from seabird guano, hydrolyzed fish powder, feather meal and blood
meal. Across temperatures, all fertilizers had equivalent N mta after 15 days, with blood meal
having a slight advantage after 2 months.
According to Shepherd, 137 blood meal is sometimes not recommended for organic gar-
dening because it can damage young tender plants in warm moist conditions.
d) Scientific use or research
Animals can produce useful medical substances in their blood, like vaccines, hormones,
and antibodies, which are important for basic research, diagnostic tests, and medical treat-
ments. Human biology is very much like that of many other animals. That is why results
from animal experiments apply to people. Most laboratory animals have the same set of
organs (heart, lungs, liver, and so on) that work in the same way as they do in humans. 138
e) Laboratory uses
Serum
It is widely used in cell culture, microbiology procedures and diagnostic kits. Blood
from donor animals continues to play an important role as a nutritional supplement in mi-
crobiological culture media. Many isolated blood components are used in chemical analysis
or as nutritive supplements. Blood plasma is used as a diluent for boar and bull semen.
134. Gagnon B., S. Berrouard (1994), "Effects of several organic fertilizers on growth of greenhouse
tomato transplants", Canadian Journal of Plant Science, 74: 167-168.
135. Ciavatta C, M. Govia, L. Sittia, C. Gessa (1997), "Influence of blood meal organic fertilizer on
soil organic matter: A laboratory study" ', Journal of Plant Nutrition, 20 (11): 1573-1591.
136. Hartz T. K., P. R. Johnstone (2006), "Nitrogen Availability from high-nitrogen-containing Or-
ganic Fertilizers", Hort Technology, 16 (1): 39-42.
137. Shepherd A. (2003), How to make soil and save Earth, CAT Publications, Machynlleth,
Powys, UK.
138. To know more about see: Giridharan N. V., Vijay Kumar, Vasantha Muthuswamy (2000), Use
of Animals in Scientific Research, Indian Council of Medical Research, Ministry of Health & Family
Welfare, New Delhi.
45
Raffaella Leoci
Factors
The 13 blood clotting factors that control clot are: Factor I = Fibrinogen, Factor II =
Prothrombin, Factor III = Tissue factor, Factor IV = Calcium, Factor V = Labile factor, Fac-
tor VI - (Does not exist as it was named initially but later on discovered not to play a part
in blood coagulation), Factor VII = Stable factor, Factor VIII = Antihemophilic factor A,
Factor IX = Antihemophilic factor B or Christmas factor, Factor X = Stuart Prower factor,
Factor XI = Antihemophilic factor C, Factor XII = Hageman factor, and Factor XIII = Fibrin
stabilizing factor.
Coagulation factors (also known as Blood Clotting Factors, Clotting Factors, or Fac-
tor Assays) or by the individual factor number (Factor I, Factor II, etc.) or name (Fi-
brinogen, Prothrombin, etc.). Coagulation factor tests may be carried out in the presence
of excessive bleeding or hematomas or a prolonged Prothrombin Time (PT) or Partial
Thromboplastin Time (PTT). These tests are used as screening tools to determine whether
there is a coagulation problem. Factor activity may be measured in case of appropriate sig-
nals of having an acquired condition that is causing bleeding, such as vitamin K deficiency
or liver disease.
Factor testing may be done when bleeding episodes begin early in life and an inherited
coagulation factor deficiency is suspected. In this case, other family members may also be
tested to help confirm the person's diagnosis.
Blood albumin
The albumin blood test measures the amount of albumin in the blood serum. 139 It can
be used to evaluate how the liver and kidneys are functioning. 140 Low levels (hypoalbumine-
mia) can indicate liver damage, while high levels usually reflect dehydration. Low albumin
levels can also be seen in inflammation, shock, malnutrition, excess excretion by the kidneys
(as in nephrotic syndrome), excess loss in bowel (protein-losing enteropathy), burns (plasma
loss in the absence of skin barrier), redistribution (hemodilution as in pregnancy), and muta-
tion causing analbuminemia (very rare), etc. 141
139. Tietz N. W. (Edt.) (1995), Clinical Guide to Laboratory Tests, 3rd ed. W. B. Saunders, Philadel-
phia, PA; Nicoll D., S. J. McPhee, M. Pignone, Chou T. M. Detmer (edts) (2001), Pocket Guide to
Diagnostic Tests, 3rd. McGraw-Hill, New York; Ballmer P. E., M. A. McNurlan, E. Milne, S. D. Heys,
V. Buchan, A. G. Calder, P. J. Garlick (1990), "Measurement of albumin synthesis in humans: a new
approach employing stable isotopes", American Journal of Physiology, 259 (6 Pt 1): E797— E803.
140. Berk P. D., K. M. Korenblat (2007), Approach to the patient with jaundice or abnormal liver test
results (in: L. Goldman and D. Ausiello, Edts. Cecil Medicine, Saunders Elsevier, Philadelphia, Pa;
Doumas B. T, W. A. Watson, H. G. (1971), "Biggs Albumin standards and the measurement of serum
albumin with bromcresol green", Clinica Chimica Acta,i\ (1): 87-96.
141. Among others: Fearon K. C, J. S. Falconer, C. Slater, D. C. McMillan, J. A. Ross, T. Preston
(1998), "Albumin synthesis rates are not decreased in hypoalbuminemic cachectic cancer patients with
an ongoing acute-phase protein response", Annals of Surgery, 227 (2): 249-254; Ballmer P. E., A. E
Ochsenbein, S. Schutz-Hofmann (1994), "Transcapillary escape rate of albumin positively correlates
with plasma albumin concentration in acute but not in chronic inflammatory disease", Metabolism, 43
(6): 697-705; Fleck A., G. Raines, F. Hawker, J. Trotter, P. I. Wallace, I. M. Ledingham, K. C. Caiman
(1985), "Increased vascular permeability: a major cause of hypoalbuminaemia in disease and injury",
Lancet, 1 (8432): 781-784.
46
Animal by-products (ABPs): origins, uses, and European regulations
Fibrinogen
It is a protein produced by the liver that helps stop bleeding by helping blood clots to
form. A blood test can be done to know how much fibrinogen there is in the blood. Normal
levels are about 1.5-3 g/1, depending on the method used. In typical circumstances, fibrino-
gen is measured in citrated plasma samples in the laboratory, however the analysis of whole
blood samples by use of thrombelastometry is also possible. Higher levels (>3.43 g/1) are
often associated with cardiovascular disease. Fibrinogen levels may elevate in any form of
inflammation, as it is an acute-phase protein. 142 In pregnancy, fibrinogen levels increase to
an average of 4.5 g/1, compared to an average of 3 g/1 in non-pregnant women.
The acute phase response is a physiologic reaction to infection or trauma. The increase
in plasma acute phase proteins provides an objective measure of inflammation in response to
disease and therapeutic intervention. 143 Research in human and veterinary medicine has fo-
cused on acute phase proteins as an early marker in inflammation that can indicate the time
and degree of symptom. The two most common acute phase proteins evaluated in horses are
fibrinogen and serum amyloid A (SAA). 144 SAA increases rapidly (more than 100 fold) after
tissue injury but has a short half-life. Fibrinogen increases more slowly (about 24-72 hours)
after the induction of inflammation but to a lower degree (among 1-10 fold).
For this reason it is used in veterinary medicine as an inflammatory marker 145 . Fibrinogen
and Haptoglobin are good markers of chronic inflammatory disease and very useful for moni-
toring out-patients in which samples are taken less frequently. 146 In horses, a Fibrinogen level
above the normal range of 1.0-4.0 g/1 suggests some degree of systemic inflammatory response.
Haptoglobin levels correlate with the clinical status of disease in cases of musculoskeletal, res-
piratory and digestive pathologies. The presence of an inflammation process was better con-
firmed by Fibrinogen presence in cases of cutaneous and musculoskeletal pathologies. When
considering a wide range of diseased horses, it appeared that Haptoglobin has a better sensi-
142. See: Hulten C, U. Gronlund, J. Hirvonen, R. -M. Tulamo, M. M. Suominen, G. Marhaug, M.
Forsberg (2002), "Dynamics in serum of the inflammatory markers serum amyloid A (SAA), hap-
toglobin, fibrinogen and a2-globulins during induced noninfectious arthritis in the horse", Equine
Veterinary Journal, 34 (7): 699—704.
143. Crisman M., W. Scarratt, K. Zimmerman (2008), "Blood proteins and inflammation in the
horse", Veterinary Clinics of North America: Equine Practice, 24 (2): 285-297.
144. Hulten C, et al., "Dynamics in serum of the inflammatory markers serum amyloid A(SAA), hap-
toglobin, fibrinogen and a2-globulins during induced noninfectious arthritis in the horse", ibid., pp.
699-704; Jacobsen S., H. Thomsen, S. Nanni (2006), "Concentrations of serum amyloid A in serum
and synovial fluid from healthy horses and horses with joint disease", American Journal of Veterinary
Research, 67 (10): 1738-1742.
145. Allen B. V., S. E. Kold (1988), "Fibrinogen response to surgical tissue trauma in the horse",
Equine Veterinary Journal, 20 (6): 441-443; Larsson A., J. Bjork, C. Lundberg (1997), "Nephelometric
determination of rat fibrinogen as a marker of inflammatory response", Veterinary Immunology and
Immunopathology, 59 (1): 163-169.
146. Paolaggi J. B., D. Chaouat, D. Barres, H. Hoffman, L. Auquier (1982), "Comparative variations
of the sedimentation rate, haptoglobin and orosomucoid in rhizomelic pseudopolyarthritis and tem-
poral arteritis. Attempt at a definition of biological parameters for monitoring these diseases", Revue
du rhumatisme et des maladies osteo-articulaires, 49 (6): 413-419.
47
Raffaella Leoci
tivity than Fibrinogen. 147 Mair and Linnenkohl 148 hypothesized that patients presenting for
colic associated with inflammatory conditions (peritonitis, colitis, inflammatory bowel disease,
strangulating surgical lesions) would display significantly higher concentrations of Fibrinogen
and SAA compared to patients that presented for non-specific medical colic.
According to Borges et al. 149 measurement of plasma iron concentration better reflected
acute inflammation than did fibrinogen concentration.
Globulins
The two main proteins in the blood are albumin and globulin both produced by the
liver. Others are made by the immune system. Globulin carries essential metals through
the bloodstream and carries them to the various parts of the body and helps to fight infec-
tions. Globulin proteins include enzymes, antibodies and more than 500 other proteins.
All plasma proteins except albumin and prealbumin are globulins. The plasma globulins are
separated into five fractions by serum protein electrophoresis (SPE). 150 In order of decreasing
electrophoretic mobility these fractions are the alphal-, alpha 2-, betal-, and beta 2-globu-
lins, and the gamma globulins.
Sphingomyelin
It is one of the major lipids of the plasma membranes of mammalian cells. It is a sphin-
golipid found in animal cell membranes, especially in the membranous myelin sheath that
surrounds nerve cell axons. Its function remained unclear until recently, when it was found
to have a role in signal transduction. 151 Analysis of sphingomyelin can be carried out by
means of chromatography 152 or by mass spectrometry. 153
147. Oukacha R, P. Bertrand, J. Bustin, M. Humblet, H. Amory, J. Godeau (2005), Relationship
between haptoglobin, fibrinogen and clinical status in horses, 5th International Colloquium on Animal
Acute Phase Proteins, Dublin, Ireland, March 14-15, Poster 13, p. 62.
148. Mair T., W Linnenkohl (201 1), The use of serum amyloid A (SAA) and fibrinogen as disease markers
in equine colic, Bell Equine Veterinary Clinic, Mereworth, Kent, UK (e-mail: wlinnenkohl@gmail.com.) .
149. Borges A. S., T. J. Divers, T. Stokol, O. Hussni Mohammed (2007), "Serum iron and plasma
fibrinogen concentrations as indicators of systemic inflammatory diseases in hones", Journal of Veteri-
nary Internal Medicine, 21 (3): 489-494.
150. For more details see: Tiselius A. (1937), "Electrophoresis of serum globulin. Electrophoretic
analysis of normal and immune sera", Biochemical Journal, 31 (9): 1464-1477; Peacock A. C, S.
L. Bunting, K. G. Queen (1965), "Serum protein electrophoresis in acrylamide gel: patterns from
normal human subjects", Science, 147 (3664): 1451-1453; Vanderschaeghe D., E. Debruyne, H. Van
Vlierberghe, N. Callewaert, J. Delanghe (2009), "Analysis of gamma-globulin mobility on routine
clinical CE equipment: exploring its molecular basis and potential clinical utility", Electrophoresis, 30
(15): 2617-23; El-Zarkouny S. Z, M. M. Shaaban, J. S. Stevenson (2011), "Blood metabolites and
hormone-based programmed breeding treatments in anovular lactating dairy cows", Journal of Dairy
Science, 94 (12): 6001-10.
151. Kolesnick R. (1994), "Signal transduction through the sphingomyelin pathway", Molecular and
chemical neuropathology, 21 (2-3): 287-97.
152. Ramstedt B., P. Leppimaki , M. Axberg, J. P. Slotte (1999), "Analysis of natural and synthetic
sphingomyelins using high-performance thin-layer chromatography", European Journal of Biochem-
istry, 266 (3): 997-1002; Lee S., Youn-Sun Lee, Kyeong-Mi Choi, Kwang-Sik Yoo, Dong-Mi Sin,
Wonkyun Kim, Yong-Moon Lee, Jin-Tae Hong, Yeo-Pyo Yun, Hwan-Soo Yoo (2012), "Quantitative
Analysis of Sphingomyelin by High-Performance Liquid Chromatography after Enzymatic Hydroly-
sis", Evidence-Based Complementary and Alternative Medicine, Article ID 396218: 1-9.
153. Sullards M. C. (2000), "Analysis of sphingomyelin, glucosylceramide, ceramide, sphingosine, and
48
Animal by-products (ABPs): origins, uses, and European regulations
Catalase
It is an extraordinary enzyme. Catalase is ubiquitous and is found in animal 154 and in
plant cells. It catalyzes the decomposition of hydrogen peroxide (H 2 0 2 ) to water and oxy-
gen. 155 This enzyme is used in laboratories as a tool for learning the effect of enzymes upon
reaction rates. The test (catalase test) is one of the main tests used by microbiologists to
identify species of bacteria. 156
Agglutination test
It is a blood test used to identify unknown antigens. Blood with the unknown antigen
is mixed with a known antibody; whether or not agglutination occurs helps to identify the
antigen (qualitative agglutination test). Agglutination tests can also be used to measure the
level of antibodies to particulate antigens. In this test, sample serial dilutions are tested for
antibody and then a fixed number of red blood cells, bacteria, or other such particulate anti-
gens are added to determine the maximum dilution for agglutination. Agglutination testing
is also used in tissue matching, blood grouping, and diagnosis of infections. 157
Diagnostic microbiology
Microbes such as bacteria, protozoans, and fungi play a role in many disease pro-
cesses. Since the 1960s, numerous ingenious innovations have been introduced and used
in clinical microbiology laboratories. Miniaturization, plastic disposables, and commercial
medium production are the norm in present-day microbiology laboratories. Mechaniza-
tion of analytical microbiological procedures have given way to automation. Robotization,
now available for the routines of chemistry, hematology, endocrinology, some serology,
and urinalysis, may eventually find applications in some, if not many, aspects of clinical
microbiology. 158
Infectious diseases are often diagnosed following cultures of samples isolated from
the infection site. Many bacteria, fungi, parasites and viruses can be grown in a lab when
appropriate conditions are used. Precise characteristics of the growing cultures can be used
to identify specific microbe. Diagnostic cultures are commonly used to identify infectious
sphingosine 1-phosphate by tandem mass spectrometry", Methods in Enzymology, 312: 32-45; Shaner
R. L, J. C. Allegood, H. Park, E. Wang, S. Kelly, C. A. Haynes, M. C. Sullards, A. H. Merrill, Jr.
(2009), "Quantitative analysis of sphingolipids for lipidomics using triple quadrupole and quadrupole
linear ion trap mass spectrometers", The Journal of Lipid Research, 50 (8): 1692—1707.
154. Murray M. R., T. J. Reid, A. Sicignano, N. Tanaka, M. G. Rossmann (1981), "Structure of beef
liver catalase" , Journal of Molecular Biology, 152 (2): 465—499.
155. Chelikani P., I. Fita, P. C. Loewen (2004), "Diversity of structures and properties among cata-
lases", Cellular and Molecular Life Sciences, 61 (2): 192-208.
156. Gagnon M., W. Hunting, W. B. Esselen (1959), "A new method for catalase determination",
Analitical Chemistry, 31 (1): 144-146; Duke P. B„ J. D. Jarvis (1972), "The catalase test — a cautionary
tale", The Journal of Medical Laboratory Technology, 29 (2): 203—204.
157. A particular use of this test is reported by: Verloo D., E. Magnus, P. Biischer (2001), "General
expression of RoTat 1.2 variable antigen type in Trypanosoma evansi isolates from different origin",
Veterinary Parasitology, 97 (3): 183-189.
158. See: Isenberg H. D. (2003), "Clinical Microbiology: Past, Present, and Future", Journal of Clinical
Microbiolology, 41 (3): 917-918.
49
Raffaella Leoci
microbes from samples isolated from urine, stool, genital tract, throat, skin, blood, and
the spinal cord. 159 ' 160
f) Pharmaceutical uses
Animals and their parts have been used as medicinal resources for the treatment and
relief of a myriad of illnesses and diseases in practically every human culture. Pliny tells us
that the blood of animals (human blood as well) was administered in his time for curative
purposes; so likewise, today the blood of the ox is in medicinal vogue in certain parts of the
Eastern Hemisphere.
Blood is an excellent source of raw materials for making pharmaceuticals and to carry
out analysis of the following: immunoglobulins, thrombin (blood clotting), factors, sutures,
fibrinogen, fibrinolysin, fibrin products, serotonin, kalikrenis, plasminogen, plasma extend-
ers, and transfusions. Chemically the blood of animals contains a considerable quantity of
iron, in addition to albumin, fibrin, hydrogen, some traces of prussic acid, and some empy-
reumatic oil. The serum, or thin part of the blood, includes sulphur. Many blood products
are made from the plasma component of blood. Plasma contains a large number of proteins,
each of which performs a different role within the blood.
Immunoglobulins are one of the most important mechanisms the body has to protect
itself against infections. If the patient has depleted quantities in his or her blood, the use of
immunoglobulin concentrates enables replacement. These blood products are widely used
for immune deficiencies, such as for cancer patients undergoing chemotherapy.
The products most commonly isolated from plasma and their uses are overviewed in
the following: 161
• Whole Blood Transfusions or broken down into components;
• Red Cells Anemia and replacement of traumatic or surgical
blood loss;
• Platelets Hemorrhage;
• Fresh Frozen Plasma Bleeding problems after trauma or liver transplants;
• Albumin Blood volume replacement i.e. after accident;
159. Isenberg H. D., A. Reichler, D. Wiseman (1971), "Prototype of a fully automated device for
determination of bacterial antibiotic susceptibility in the clinical laboratory", Applied Microbiology, 22
(6): 980-986.
160. To be informed of the latest developments in clinical microbiology and the diagnosis and treatment
of infectious diseases, among many, see: Mahon C. R., D. C. Lehman, G. Manuselis, Jr. (2007), Textbook
of Diagnostic Microbiology, Volume 355, Elsevier Science - Health Science Division, p.1211; Diagnostic
Microbiology and Infectious Disease 2012, Elsevier, Edt. Ronald N. Jones, 74 (1): 323-434; Forbes B. A.,
D. F. Sahm, A. S. Weissfeld (2007), Bailey & Scott's Diagnostic Microbiology, Elsevier Science.
161. The 2009 Report on Blood and Blood Derivatives and Fractions excluding those used for passive
immunization and diagnostics: World Market Segmentation by City. Parker, Philip M.//City Segmen-
tation Reports; 1/3/2010, pN. PAG.
50
Animal by-products (ABPs): origins, uses, and European regulations
Immunoglobuline
Factor VIII
Factor VIII/vWF
(von Willebrand complex)
Factor IX
Factor XI
Alpha- 1 -pro teinaise inhibitor
C-l -esterase inhibitor
Protein C
Prothrombin Complex
Concentr.
Anticoagulation Complex
Antithrombin III
Fibrinogen
Thrombin
Immune deficiencies i.e. often used with patients with
Cancer receiving chemotherapy;
Hemophilia A;
von Willebrand Disease;
Hemophilia B;
Hemophilia A
Acquired and congenital emphysema. Cystic fibrosis;
Hereditary angioedema. Heart attacks;
Purpura fulminans. Couramin induced skin necrosis;
Bleeding disorders. Hemophilia B;
Bleeding disorders;
Deep Vein Thrombosis. Pulmonary embolus;
Stopping of bleeding or hemorrhaging in surgery;
Stopping of bleeding or hemorrhaging in surgery.
Brand and coll., 162 Nyberg and coll., 163 and Liebmann and coll. 164 have studied the proper-
ties of opioid peptides derived from hemoglobin.
Immunoglobulins
Glycoprotein molecules that are produced by plasma cells in response to an immunogen
function as antibodies, are called immunoglobulins. These immunoglobulins are synthesized
by B-lymphocytes. Their derivative plasma cells are found in the blood serum and in other
body fluids and tissues, including the urine, spinal fluid, lymph nodes and spleen. Immuno-
globulin molecules consist of two kinds of polypeptide chains: heavy chains (H-chains) and
light chains (L-chains). There are five antigenically different kinds of H-chains, designated
y, |i, a, 5 and e, based on differences in the amino acid sequences in the constant region of
the H-chains. This difference is the basis for the classification of immunoglobulins. Classes
vary in their chemical structure and in the number of antigen-binding sites. The classes of
immunoglobulins can divided into subclasses based on small differences in the amino acid
sequences in the constant region of the heavy chains.
162. Brand V., C. Gramsch, F. Lottspeich, R. Mertz, K. H. Jaeger, A. Herz (1986), "Novel opioid
peptides derived from hemoglobin: hemorphins", European Journal of Pharmacology, 125 (2): 309-10.
163. Nyberg R, K. Sanderson, E. L. Glamsta (1997), "The hemorphins: a new class of opioid peptides
derived from the blood protein haemoglobin", Biopolymers, 43 (2): 147—56.
164. Liebmann C, U. Schrader, V. Brand (1989), "Opioid receptor affinities of the blood-derived
tetrapeptides hemorphin and cytochrophin", European Journal of Pharmacology, 166 (3): 523-6.
51
Raffaella Leoci
The five classes of immunoglobulins (Ig) are: IgA, 165 IgD, 166 IgE, 167 IgG, 168 IgM. 169 Only
IgG, IgM and IgA and are present in all species of domestic animals. 170
Blood clotting or blood clot
It consists of a plug of platelets enmeshed in a network of insoluble fibrin molecules.
When blood vessels are cut or damaged, the loss of blood from the system must be stopped
before shock and possible death occur. This is accomplished by solidification of the blood,
a process called coagulation or clotting. 171 Blood clots are healthy and lifesaving when they
165. IgA is present in low concentrations in the serum, but it is the major immunoglobulin of secre-
tions and has a major first-line defense role in infections. Two IgA molecules are linked by a polypep-
tide called the secretory piece and by a J chain. Secretory IgA is present in nonvascular fluids, such as
saliva, bile, synovial fluid, and intestinal and respiratory tract secretions. Both secreted and circulating
IgA types are known to have antiviral properties; their production is preferentially stimulated by local
administration of antigens such as oral and aerosol immunizations.
166. IgD is found in trace quantities in the serum in humans and chickens. Its function is uncertain.
167. IgE is present in low levels in serum and is generally present in increased levels in individuals with
allergy. It has not been found in the chicken. Following exposure to antigen (allergen), and its IgE
binding to the Fab of two adjacent molecules, perturbations of the cell membrane are produced, lead-
ing to the release of vasoactive amines (histamine, serotonin, etc.) which are the mediators of anaphy-
laxis and atopic reactions, including urticaria, asthma, hayfever and gastroenteritis. Allergic reactions
(urticaria, atopy, anaphylactic shock, etc.) are examples of IgE-mediated reactions. In humans and dogs
there is an inherited predisposition for certain individuals to produce IgE.
168. IgG is the most abundant of the five classes of immunoglobulins. It is the only that crosses the
placenta and is the major component of passive maternal antibody transfer via colostrum and yolk. It
is the major antibody in the secondary humoral response of immunity.
169. IgM is the first antibody produced in the primary immune response. Like the IgG, IgM bound to
antigen activates the complement system. These two immunoglobulins are specific antitoxins against
the toxins of diphtheria, tetanus, snake venoms, botulism and anthrax microorganisms, and are used
in defence against most infectious diseases.
170. For more information, see among others: Fahey J. L. (1965), "Antibodies and Immunoglobu-
linsll. Normal Development and Changes in Disease", The Journal of American Medical Association
(JAMA), 194 (3): 255-258; Johansson S. G. O., M. B. Uppsala (1967), "Raised levels of a new immu-
noglobulin Class (Ignd) in Asthma", The Lancet, 290 (7523): 951-953; Butler J. E. (1969), "Bovine
Immunoglobulins: A Review" , Journal of Dairy Science, 52 (12): 1895-1909; Kalpaktsoglou P. K., R.
Hong, R. A. Good (1973), "The five classes of immunoglobulins in normal C3H and BALB/c mice",
Immunology, 24 (2): 303—314; Ogra S. S., P. L. Ogra (1978), "Immunologic aspects of human co-
lostrum and milk: I. Distribution characteristics and concentrations of immunoglobulins at different
times after the onset of lactation", The Journal of Pediatrics, 92 (4): 546-549; Rose M. E., E. Orlans
(1981), "Immunoglobulins in the egg, embryo and young chick", Developmental & Comparative Im-
munology, 5 (1): 15-20; Hombach J., T. Tsubata, L. Leclercq, H. Stappert, M. Reth (1990), "Mo-
lecular components of the B-cell antigen receptor complex of the IgM class", Nature, 343: 760 — 762;
Boulinier T, V. Staszewski (2008), "Maternal transfer of antibodies: raising immuno-ecology issues",
Trends in Ecology & Evolution, 23 (5): 282-288; Schroeder H. W. Jr., L. Cavacini (2010), "Structure
and function of immunoglobulins", Journal of Allergy and Clinical Immunology, 125 (2): S41-S50.
171. Miller J. L. (1996), Blood Coagulation and Fibrinolysis, Clinical Diagnosis and Management by
Laboratory Methods, edt. by J. B. Henry, 19th ed., W. B. Saunders Co., Philadelphia; Hardaway R.
M., C. H. Williams (1996), "Disseminated Intravascular Coagulation: An Update", Comprehensive
Therapy, 22 (11): 737-743.
52
Animal by-products (ABPs): origins, uses, and European regulations
stop unwanted bleeding. However harmful blood clots can also form, causing serious dam-
age. Most heart attacks and strokes result from the sudden formation of a blood clot on a
cholesterol plaque inside an artery in the heart or brain. The process of blood clotting is trig-
gered whenever the normal flow of blood is exposed to certain substances; there are a lot of
different substances, called thrombogenic substances that promote formation of thrombus
(another name for a clot). These substances are located in the skin or in blood vessel walls.
Examples of these thrombogenic substances are tissue factor, collagen, and von Willebrand
factor. While the formation of clot is essential to life and can be regulated, inappropriate clot
formation, especially in the brain or lungs, can be life-threatening.
There are many substances that inactivate the thrombin. Antithrombin III is one of
these. It is a plasma protein (a serpin) that inhibits the formation of thrombin. Heparin is
another inhibitor. It is a mixture of polysaccharides that bind to antithrombin III, inducing
an allosteric change that greatly enhances repression of thrombin synthesis.
Fibrinolysin
It is an enzyme derived from bovine plasmin or extracted from cultures of certain bac-
teria. It is used in commercial preparations only and exclusively together with the enzyme
desoxyribonuclease (extracted from bovine pancreas). Fibrinolysin and desoxyribonuclease
both act as lytic enzymes. Fibrinolysin is thought to break down fibrin in necrotic material,
while desoxyribonuclease is thought to degrade DNA residue of necrotic cells. 172 No long-
term clinical benefit was demonstrated in reducing purulent exudates or necrotic tissue. 173
Serotonin 174
It is found mainly in the gastrointestinal tract of animals. About 80 to 90% of the hu-
man body's total serotonin is located in the enterochromaffin cells in the gut, where it is used
to regulate intestinal movements. Serptonin is a monoamine neurotransmitter that regulates
mainly metabolism, mood, 175 and appetite and appears to be involved in control of sleep,
memory and learning, temperature regulation, behavior (including hallucinogenic behavior
and sexual), cardiovascular function, muscle contraction, endocrine regulation, and depres-
sion. Almost all the body's serotonin is used by muscles and blood vessels. It is synthesized
in brain neurons, from the amino acid L-tryptophan, and is stored in vesicles. Serotonin
is found not only in animals, but also in fungi and plants, including fruits and vegetables.
172. Westerhof W„ F. C. Jansen, F. S. De Wit, R. H. Cormane (1987), "Controlled double-blind trial
of fibrinolysin —desoxyribonuclease (Elase) solution in patient with chronic leg ulcers who are treated
before autoluogos skin grafting", Journal of American Acade Dermatology, 17 (1): 32-39
173. Shai A., H. I. Maibach (2005), Wound Healing and Ulcers of the Skin: Diagnosis and Therapy - The
Practical Approach, Spring- Verlag, Berlin-Heidelberg-New York, p. 128.
174. Miiller C. P., B. L. Jacobs, Edtr. (2010), Handbook of the Behavioral Neurobiology of Serotonin,
Volume 21, Elsevier B.V., pp. 3-818.
175. Animal research suggests that central serotonergic neurons are involved in behavioral suppres-
sion, particularly anxiety-related inhibition. The hypothesis linking decreased serotonin transmission
to reduced anxiety as the mechanism in the anxiolytic activity of benzodiazepines conflicts with most
clinical observations (See: Soubrie P. (1986), "Reconciling the role of central serotonin neurons in hu-
man and animal behaviour", Behavioral and Brain Sciences, 9: 319-335).
53
Raffaella Leoci
Shi Wu Wen and others 176 have studied the safety of the use of selective serotonin reup-
take inhibitors in pregnancy. They claim that the risks of low birth weight, preterm birth, fe-
tal death, and seizures were increased in infants who were born to mothers who had received
selective serotonin reuptake inhibitor therapy.
Kallikreins
They are enzymes capable of cleaving peptide bonds in proteins. They are a subgroup
of serine proteases. Any of two groups of serine endopeptidases that are widely distributed
in mammalian tissues and body fluids, including blood. In 1934, Werle 177 reported finding
great amounts of the substances in the pancreas of human men and other animals, such that
the pancreas could be taken for its site of origin. He named it kallikrein from the Greek word
for pancreas. Later, Werle et al. 178 identified kallikrein as a proteolytic enzyme ('ferment')
that liberates the biologically highly active basic polypeptide 'DK', or kallidin (i.e lys-brady-
kinin) from a blood plasma protein called kallidinogen or kininogen (H- and L-kininogen).
Human tissue kallikrein-related peptidases (KLKs) are a family of 15 serine proteases
with diverse physiological functions. The vital importance of the kallikrein-kinin system
for basic mechanisms in biochemistry, patho/physiology, pharmacology and more recently,
molecular biology and cell biology, and the great interest and practical benefit of the system
to clinical medicine, has stimulated scientists from various disciplines worldwide to become
involved in kallikrein-kinin research. KLKs play important roles in different physiologic
processes, such as regulation of cell growth and differentiation, tissue remodeling, skin des-
quamation, and human semen liquefaction, 179 etc. Several members of the human tissue
kallikrein-related peptidase (KLK) family are emerging cancer biomarkers. The aim of this
study was to analyse the expression of a panel of KLKs in colorectal cancer and to find out if
the multiparametric combination of them can increase the accuracy of prediction of patients
survival beyond the traditional clinical information. 180
Plasmin
Like trypsin, it belongs to the family of serine proteases. Plasmin is an enzyme present
in blood that degrades many blood plasma proteins, particularly, fibrin clots. The generation
of plasmin by plasminogen activators is a physiologic process in animals that dissolves blood
clots and promotes wound healing, blood vessel growth, and the migration of normal and
176. Wen S. W., Q. Yang, P. Garner, W. Fraser, O. Olatunbosun, C. Nimrod, M. Walker (2006), "Se-
lective serotonin reuptake inhibitors and adverse pregnancy outcomes", American Journal of Obstetrics
and Gynecology, 194 (4): 961-966.
177. Werle E. (1934), "Zur Kenntnis des Haushalts des Kallikreins", Biochemische Zeitschrift, 269:
415-434.
178. Werle E., W. Gotze, A. Kappler (1937), "Uber die Wirkung des Kallikreins auf den isolierten
Darm und iiber eine neue darmkontrahierende Substanz", Biochemische Zeitschrift, 289: 217-233.
179. Pampalakis G., G. Sotiropoulou (2007), "Tissue kallikrein proteolytic cascade pathways in nor-
mal physiology and cancer", Biochimica et Biophysica Acta, 1776 (1): 22-31.
180. Talieri M., L. Li, Y. Zheng, D. K. Alexopoulou, A. Soosaipillai, A. Scorilas, D. Xynopoulos, E. P.
Diamandis (2009), "The use of kallikrein-related peptidases as adjuvant prognostic markers in colorec-
tal cancer", British Journal of Cancer, 100 (10): 1659-1665.
54
Animal by-products (ABPs): origins, uses, and European regulations
cancerous cells. 181 Streptokinase and urokinase have been the standard agents available for
many years, but in recent years the most exciting change in the field has been the develop-
ment of a new generation of plasminogen activators, the principal one being a tissue plasmi-
nogen activator. 182 A tissue plasminogen activator is a clot-dissolving enzyme that is naturally
produced by cells in the walls of blood vessels; it catalyzes the conversion of plasminogen to
plasmin. Plasmin is the main proteolytic enzyme in milk and has been found to be associated
with enhanced casein hydrolysation. 183
The role of the plasminogen-plasmin system in fertilization is unknown, although its
dysfunction has been associated with subfertility in humans. Coy and coll. 184 have recently
detected and quantified plasminogen in the oviductal fluid of two mammals and showed a
reduction in sperm penetration during IVF when plasminogen is present. The objective of
this study was to describe the mechanism by which the plasminogen-plasmin system regu-
lates sperm entry into the oocyte.
According to Marder and co-workers, 185 plasmin is tolerated without bleeding at a
several-fold higher amount than that needed for thrombolysis, which is in contradistinc-
tion to plasminogen activators that risk bleeding at any effective thrombolytic dose. Plas-
min has been safe in a current trial in patients with peripheral arterial or graft occlusion,
and efforts are now directed toward use in therapy of stroke caused by cerebral artery
occlusion. 186
Saaand coll. 187 have undertaken a study to examine the effect of plasmin on sperm
viability and sperm— oocyte interaction during in vitro fertilization in the pig. The results
suggest that plasmin might play a role in events related to fertilization.
g) Veterinary biological uses (also called veterinary biologies) are those used for the
diagnosis and treatment of diseases in animals including vaccines, bacterins,
antitoxins, toxoids, immunomodulators.
181. Gladysheva I. P., R. B. Turner, I. Y. Sazonova, L. Liu, G. L. Reed (2003), "Coevolutionary pat-
terns in plasminogen activation", PNAS, 100 (16): 9168-9172.
182. Hirsh J., A. G. Turpie (1990), "Use of plasminogen activators in venous thrombosis", World
Journal of Surgery, 14 (5): 688-93.
183. Battacone G., E. A. Cannas, A. Mazzette, C. Dimauro, G. Enne (2005), "Why does the increase
of plasmin worsen the coagulation properties of milk in dairy sheep?", Italian Journal of Animal Sci-
ence, 4 (Suppl. 2): 342-344.
184. Coy P., M. Jimenez-Movilla, F. A. Garcia- Vazquez, I. Mondejar, L. Grullon, R. Romar (2012),
"Oocytes use the plasminogen-plasmin system to remove supernumerary spermatozoa", Oxford Jour-
nals - Human Reproduction, 27 (7): 1985-1993.
185. Marder V. J., R. Jahan, T. Gruber, A. Goyal, V. Arora (2010), "Approaches to Thrombolysis.
Thrombolysis With Plasmin. Implications for Stroke Treatment", Stroke, 41: S45-S49.
186. See also: Riviere J. E., M. G. Papich (2009), Veterinary Pharmacology & Therapeutics, Wiley-
Blackwell, 9 th Edition, pp. 685-6.
187. Saa S. J., H. H. Rheeb, H. T. Cheonga, B. K. Yanga, C. K. Park (2006), "Effects of plasmin on
sperm-oocyte interactions during in vitro fertilization in the pig", Animal Reproduction Science, 95
(3-4): 273-282.
55
Raffaella Leoci
h) Industrial uses
Adhesives
They may be defined as any substance capable of attaching materials together by means
of surface attachment. Knowledge and use of adhesives is very old. The ancient Egyptians,
3500 years before Christ, knew well the art of veneering and used adhesives to attach deco-
rations to wood. Clay, mud, and dung, along with mixtures of these substances, must also
be regarded as adhesives and have been used for centuries. Until the Second World War,
essentially all of the glues were of natural origin. Adhesives based on synthetic polymers
were introduced just before that war and now surpass most of the older natural glues in
importance for wood bonding. 188
Fish skin, bones, horns and hides of cows, seal brain and blood are a little part of the list
of animals used for manufacturing glues. As a consequence, animal glue includes hide glue,
bone glue, fish glue, blood glue and skin glue. They are generally classified into three main
types: those made from hide and bone, those made from fish skins, and those made from
fresh or dried beef blood. 189 Hide glue is used in woodworking. Besides horses, hide glue uses
cows' hooves, bones, and hides.
Blood glues are either made from fresh animal blood or soluble dried beef blood, a by-
product of the meat packaging operation. The latter is produced by evaporating the serum
from fresh whole blood. The adhesive can be made from any animal blood, such as bovine,
porcine, or avian blood.
Water-resistant glues based on blood albumen are made by mixing the dried blood
powder, obtained from cattle or hog slaughter houses, with water which is then activated by
the addition of an alkali such as slaked lime and caustic soda and perhaps other chemicals
such as ashes, and/or alum.
Another type of adhesive is made by adding an anticoagulant and a preservative to fresh,
whole animal blood without dewatering the blood. Lime is then added to the anti-coagu-
lated, preserved blood, and the pH is adjusted to 9-11. A curing agent (such as potassium
silicate, sodium silicate and aluminum dihydrogen phosphate) and ammonia are then added
to the mixture to yield the animal blood adhesive. 190
Blood glues have been used as plywood adhesive, as paint for brickwork, as adhesive for
filling joints between brick and building stones, and in food packaging since the bonds are
odorless, nontoxic, and tasteless. 191 A typical use was the bonding of cork disks in metal bottle
caps. In some countries, blood glues made of cow's blood are used for plywood manufacture.
Animal blood can also be used to make water-soluble ketone-aldheyde and phenol-
188. The introduction of synthetic resin glues proved more convenient to use and some of them ex-
celled the blood albumin glues in durability under severe service conditions.
189. Blood glues may be made either from the fresh blood of slaughtered animals or from the dried
soluble blood albumin. To make the use of fresh blood feasible, the supply must be readily accessible
to the place of manufacture, inasmuch as rapid decomposition takes place and renders it unsatisfactory
for its purpose. Alternatively you can use the fresh blood treated with a preservative.
190. Gunasekaran S., H. Lin, Glue from slaughterhouse animal blood, United States Patent Application
20100018436.
191. See: Hubbard J. (1977), Animal Glues in Handbook of Adhesives, I. Skeist (ed.), Van Nostrand
Reinhold, New York, pp. 172-180.
56
Animal by-products (ABPs): origins, uses, and European regulations
ketone- aldheyde resin binders become more extendable and more sprayable. 192
These glues have quite good water resistant qualities but under damp conditions are
highly susceptible to mold growth and attack by bacteria.
The use of dural sealants has become common in neurosurgery. One of these is rep-
resented by a bovine albumin-glutaraldehyde combination (BioGlue 193 ). Klimo and al.,
however, have discovered that ten patients implanted with that adhesive have had wound
complications. 194
A two-component sealant composed of bovine serum albumin and glutaraldehyde (Bio-
Glue) is used to treat aortic dissections. However, some studies 195 show that polymerized
BioGlue releases amounts of glutaraldehyde that are capable of inducing cytotoxic effects
both in vitro and in vivo. Use of BioGlue should be restricted to the aortic dissection proce-
dure, as other tissues are sensitive to the amounts of glutaraldehyde released from the glue.
Research 196 conducted in the USA confirm that autologous fibrinogen, derived by poly-
ethylene glycol precipitation from the blood of an individual patient would avoid the risk of
transmitting hepatitis, and it has been shown to be relatively safe in animal studies. Fibrino-
gen-based adhesive, derived from pooled human plasma, has been used in Europe with great
success in otologic surgery, but has not been approved for use in the U.S.
The above reports and others suggest that fibrin glue can be used to seal porous vascular
grafts prior to insertion. Gundry and Behrendt 197 have studied that ability comparing blood
loss from and handling characteristics of grafts pretreated either with fibrin glue, albumin
autoclaving, or blood preclotting. They concluded that "fibrin glue or albumin is superior to
blood for pretreatment of woven grafts in limiting blood loss. Fibrin glue imparts superior
handling characteristics".
Experiments with medical glues suggest that one-third of all wounds may be "stitched"
with glues in the next few years. The adhesive have the potential to be used in tens of thousands
of operations in which the delicacy of the surgery rules out the use of sutures and staples.
For many applications natural adhesives have been replaced by synthetics; however,
animal glues, starches, gums, natural rubber cements, bitumens, and cellulose continue to
be used in large volumes. Modern synthetic adhesives do not require animal lives, and offer
192. See for example: Blood albumin glues: their manufacture, preparation and application, US Depart-
ment of Agriculture. Forest Service. Report n. 281-2, 1955, Wisconsin- USA; Campbell C. C, Cher-
ry Hill (1969), Acetone-Formaldehyde and Phenol-Acetone-Formaldheyde extended with animal blood or
soy flour and oil-in-water emulsion by, US Patent n. 3,471,420 of October 7, 1969.
193. BioGlue is used to help seal leaks around sutures (surgical stitches) or staples in large blood vessels
such as the aorta or the femoral and carotid arteries. Product name is CryoLife BioGlue Surgical Adhesive.
194. Klimo P. Jr., A. Khalil, J. R. Slotkin, E. R. Smith, M. R. Scott, L. Goumnerova (2007), "Wound
Complications Associated With the Use of Bovine Serum Albumin-Glutaraldehyde Surgical Adhesive
in Pediatric Patients", Neurosurgery, 60 (4): 305-309.
195. Fiirst W, A. Banerjee (2005), "Release of Glutaraldehyde From an Albumin-Glutaraldehyde
Tissue Adhesive Causes Significant In Vitro and In Vivo Toxicity", The Annals of Thoracic Surgery, 79:
1522-1528.
196. Weisman R. A., A. J. Torsiglieri, A. D. Schreiber, G. H. Epstein (1987), "Biochemical character-
ization of autologous fibrinogen adhesive", The Laryngoscope, 97 (10): 1186-1190.
197. Gundry S. R., D. M. Behrendt (1987), "A Quantitative and Qualitative Comparison of Fibrin
Glue, Albumin, and Blood as Agents to Pretreat Porous Vascular Grafts" ', Journal of Surgical Research,
43(1): 75-77.
57
Raffaella Leoci
several other advantages: longer shelf lives, lower freight costs, reliability and flexibility of
application. In addition, synthetic adhesives find applications in the textiles, paper, dyeing,
printing, furniture, plumbing, shoe, book, building, and automobile sectors. Many com-
munities should be using them in order not to violate their religious and moral principles.
For example, in India animal glue is still used in the manufacture of matches, books, tex-
tiles, cycles, and sports goods. The traditional Indian forehead "Bindi", the emery nail file,
sports racquet grips, and even shoes termed as "non-leather" are sometimes glued using
parts of a cow.
The United States is a major producer and consumer of glued-wood products and there-
fore of adhesive resin solids. 198 To manufacture about 60 million cubic meters/year, the adhe-
sive resin solids required to bond glued-wood products are estimated to be about 1.5 million
metric t/year. Nearly 60% of adhesive consumptions are UF (urea-formaldehyde), about
30% are PF (phenol-formaldehyde) and RF (resorcinol-formaldehyde), and the remaining
10% consist of several products, including PMDI (diphenyl methylene diisocyanate).
Increasing oil prices are boosting resin prices. Adhesive are a vital part of glued-wood
composites, their cost range contributing about 30% of the finished product. As a conse-
quence research continues to seek natural alternatives that can effectively replace synthetic
resin adhesives as a wood binder.
Casein is perhaps the first structural adhesives, and it is still in use. Production volume is
small (under 5000 t/year). Most is modified with soy flour. Soybean glues and soy-modified
casein adhesives have a long history, and, because of the green movement in costruction,
there is renewed interest in soy flour and soy protein isolates for wood-based and agri-fiber
composites. Research continues on soy products for finger jointing adhesives, as replace-
ments for animal blood in plywood production, for PF and PMDI resin blends in OSB, and
as a wheat/soy blends for plywood glue extenders. In North America, soluble animal blood
is now rare as an adhesive, but it is indispensable in PF foam adhesives for industrial and
construction plywoods. Eleven mills in North America and one in Europe were using foam
glues in 1999.
Finishes for leather and textiles
Ox blood is used as a non-thermoplastic binder in finish preparations. Added to black
or dark coloured leather, it improves the leather's depth of colour and brilliance.
The blood's fibrin fibers appear to stretch farther than any other natural fiber, up to six
times their length, before breaking according to an expert's report in 2006. 199
According to Miyata and Taira 200 collagen is a typical biological macromolecule hav-
ing been often utilized as a material with properties similar to cellulose. Nowadays its
application is becoming more widespread, ranging from classical applications such as in
the leather, gelatin, and food industries to more current uses in the biomaterial and bio-
technological fields.
198. Orr L. (2007), Wood Adhesives. A market opportunity study, USB, Midland (Mi - USA).
199. Liu W., L. M. Jawerth, E. A. Sparks, M. R. Falvo, R. R. Hantgan, R. Superfine, S. T. Lord, M.
Guthold (2006), "Fibrin Fibers Have Extraordinary Extensibility and Elasticity", Science, 313 (5787):
684-687.
200. Miyata T., T. Taira (1992), "Collagen engineering for biomaterial use", Clinical Materials, 9 (3—
4): 139-148.
58
Animal by-products (ABPs): origins, uses, and European regulations
Foam fire extinguisher 201
In about 200 BC, the Roman Ctesibius of Alexandria is credited with inventing a hand
operated fire pump able to deliver a stream of water to a fire. It was the precursor for other
variants of forcing water out of a container.
Subsequently the water inside the fire extinguisher was replaced with foam capable of
controlling flammable solid fires (substances such as wood, paper, hay, etc.), and fires fueled
by flammable oils, petrol, diesel, spirits etc. A foam fire extinguishers now contains chemical
foam, but in the past it used to contain ox blood. Ox blood inside the extinguisher reacted
with C0 2 to create a foam that, under pressure, was released to form a sealing cover over a
liquid fire. The heat from the fire would "cook" the foam forming a blanket on the surface
of the burning liquid or solid; this ox blood blanket cooled and smothered the fire in such a
way that no oxygen or heat was available to re— ignite the fire.
When protein compounds are prepared by the chemically treating various protein mate-
rials are then suitably diluted with water, they produce stable foam for fire fighting purposes.
The basic materials used in the manufacture of protein fire-fighting compounds are horn and
hoof meal, and animal blood. During the course of manufacture of horn and hoof meal a
certain amount of acid is used. In earlier times the acid was sulphuric. Compounds made by
this process tended to the formation of sludge, consisting mainly of crystals of calcium and
sodium sulphate. This compound, referred as a "sulphate compound", is no longer manufac-
tured. To overcome the sludging problem, hydrochloric acid was used and this compound is
known as "chloride compound".
A foam fire extinguisher must be non toxic, non damaging to most materials, must
extinguish fires progressively, and must prevent the re - ignition of flammable liquid fires.
Since 2003, University of Alberta (USA) scientists have been working with the agri-
culture industry to transform protein from the blood of slaughtered cows into a new fire
fighting foam. The impetus for the research was the 2003 Mad Cow crisis, which adversely
impacted many cattle-by product markets. Prior to that crisis, blood meal had been sold
as an additive to livestock feed. After the crisis, chemical companies identified a potential
market application for a protein-based foaming agent. Blood meal is high in protein that
stabilizes foam. Foam extinguishes fires. The research grew from there.
Porous concrete 202
Concrete is a strong, hard, building material composed of sand, gravel, cement, and
water. Concrete additives have been used since Roman and Egyptian times, when it was
discovered that adding volcanic ash to the mix allowed it to set-up under water. Similarly,
the Romans knew that adding horse hair made concrete less liable to crack while it hardened
201. Ratzer A. F. (1956), "History and Development of Foam as a Fire Extinguishing Medium", In-
dustrial & Engineering Chemistry, 48 (11): 2013-2016; Rivkind L. E., I. Myerson (1956), "Foams for
Industrial Fire Protection", Industrial & Engineering Chemistry, vol. 48 (11): 2017—2020; Tuve R. L.,
H. B. Peterson (1956), "Characterization of Foams for Fire Extinguishment", Industrial & Engineering
Chemistry,. 48 (11): 2024-2030; Perri J. M., C. Conway (1956), "Foam as a Fire Exposure Protection
Medium — Evaluating Effectiveness of Wetting and Protein Agents", Industrial & Engineering Chemis-
try, 48 (11): 2021-2023.
202. Sedgwick J. (1988), "Strong but sensitive", Atlantic Monthly 1991-4, 267 (4): 70-82; Weisburd S.
(1988), "Hard Science", Science news, 134 (2): 24-26.
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and adding blood to strengthened it against frost. Concrete is a porous material and add-
ing larger pored blood to the mix allows moisture to expand into the blood cells deterring
concrete cracks when temperature drop. Today, though the same ancient principles apply,
polymer fibers are used instead of horse hair and specialist "air-entraining agents" are used
instead of blood.
Plastic and cosmetic base formulations
The use of cosmetics, since ancient times, has been widespread in the societies that want
to improve appearance (e.g., by highlighting certain features of the face and/or accentuating
natural colors) or to hide imperfections of the skin. Cosmetics usually refer to substances or
preparations intended for contact with the epidermis of the human body in order to provide
functions such as cleansing, protecting, changing appearance, perfuming, altering the odor,
and improving or maintaining skin conditions. More specific functions for cosmetics are the
following: cellulite reduction, edema reduction, sebum removal, exfoliation/peeling, anti-
aging, anti- wrinkle-pucker, anti-acne-zits-pimples/spots, moisturizing or lubricating of skin,
anti-clogging of pores, coloring/tanning. 203
Many raw materials with high protein content (e.g. proteins from wheat, soybean, sun-
flower seed, peanut, rapeseed, etc., fish or meat proteins, seric albumin or egg, casein, blood,
collagen, etc.) can be used to manufacture different products, with different functional prop-
erties (adhesive, thermoplastic, elastomeric, thermoset, cosmetic, etc.). There is considerable
potential for modulating the properties of protein based materials because of the variability
of their amino-acid composition. Products made from proteins are usually biodegradable
and sometimes even edible when food-grade components and manufacturing processes are
used. 204 Cosmetics can contain albumin, a protein component of blood. Animal byproducts,
such as blood can also be used to formulate cosmetics.
In 1964, a particular cosmetic ("Magic Secret") was introduced into the US market.
The product was advertised as a wrinkle-smoothing lotion that "smooths away wrinkles in
minutes, keeps them away for hours". Magic Secret was followed by many similar cosmetic
creams, with the same features. All of these products were based on the bovine serum albu-
min extracted from cattle blood. One of the advantages of using blood serum as an albumin
source was that it was very easy to extract.
The albumin is water soluble and light in color; its concentration (albumin content) is
about 80-95%. Bovine albumin is available from several sources in three forms: as a 15%
sterile solution ready for immediate use without dilution; as a 30% solution to be diluted
with an equal volume of water prior to use, and also in the form of a freeze-dried powder
which is reconstituted with water before use.
203. De Navarre M. G. (1975), The chemistry and manufacture of cosmetics, (2nd. ed., Vols. I-IV),
Continental Press Inc., Orlando (PA - USA).
204. Guilbert S., Marie-H. Morel, N. Gontard, B. Cuq (2006), Protein-Based Plastics and Composites
as Smart Green Materials, ACS Symposium Series, vol. 921, Chapter 24, pp 334-350.
60
Animal by-products (ABPs): origins, uses, and European regulations
4.3 Bones
Bone material is a mixture of calcium phosphate and calcium carbonate with other
minerals present. The calcium lies in the form of a mixture of hydroxyapatite (HAP) crys-
tals 205 and amorphous calcium phosphate. The largest fraction of hard bone is found in the
high-density leg bones of cattle, sheep, and goats. Poultry bones are essentially all soft bones.
A natural form of HAP has applications as an absorbent, a catalyst, a dental substrate,
and as a bone substitute. Specially processed cartilage from the breast-bone of young cattle is
used by plastic surgeons to replace facial bones in human beings. In most instances, the hard
animal bones are ground into bone-protein meal and used as a source of calcium.
HAP is a crystalline material that is water insoluble. Its chemical properties and physical
structure make HAP useful as chromatographic packing material, catalyst, 206 catalyst support,
enzyme immobilization substrate, and as a component for artificial bone reconstruction.
Catalyst
The calcium in HAP can be replaced by other metals such as strontium, copper, lead,
platinum, iron, barium, etc. The Ca/P ratio and the ability to substitute other metals for
part of the calcium are the properties of HAP that influence its catalytic performance. In the
scientific literature there are a lot of reports about catalysts based on HAP with partial sub-
stitution of the calcium by platinum, copper, cobalt, nickel, etc. The applications of HAP as
catalysts include HAP with Pt, Co, Cu, Fe, Ni useful as exhaust gas catalyst to remove NOx,
C0 2 and Hydrocarbons; HAP and acidic HAP useful for partial oxidation of methane to
CO and H 2 (to use for fuel cells) or to produce formaldehyde from methanol; for amination
of alcohols; for dehydrogenation of propane to propylene; for production of ethylene from
methane (HAP with Pb); and for conversion of aromatics to alkyl aromatics (HAP with Zn,
Ni, CuCl 2 ).
The market for solid catalyst car exhausts and fuel cells is a high value-added area and
seems set for future growth on the back of exponential growth in the nanotechnology sector.
There is prior art in the use of synthetic HAP as a catalyst support.
Application for biological implant 107
Ceramics used for restoration of teeth and bones are often formed by a mixture of whit-
lockite 208 and HAP. One method of using HAP is to form a polymer-mineral composite by
precipitating the HAP onto polymer fibers. The use of polymer fibers allows formation of
the shape needed for the prosthesis. It follows the insertion of HAP onto the polymer fibers
and insert the prosthesis. In the course of time the polymer is absorbed and the HAP forms
a matrix for the natural bone to grow.
205. HAP is a crystalline mineral that has the nominal formula Ca 10 (PO 4 ) 6 (OH) , with a stoichio-
metric (Ca/P) ratio of 1,67. It is prepared reacting Ca(N0 3 ) 2 .4H 2 0 and (NH 4 ) 2 HP0 4 . The solids
obtained are ground to the desired size for the catalyst application.
206. Catalysts are at the heart of the petrochemical industry. Their sales in refining are about 2,5 billion
dollars/year, with a 2% growth rate.
207. An implant is a medical device manufactured to replace a missing or support a damaged biological
structure, or enhance an existing biological structure.
208. Whitlockite is a form of crystalline calcium phosphate with formula Ca 9 (MgFe)(P0 4 )6P0 3 OH.
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Raffaella Leoci
It is very difficult to develop the medical market for reconstructive bone and dental
applications. Perhaps non-human markets for HAP ceramics and catalysts could advance
health impact concerning human contact.
If HAP could fullfill even a small application in the ceramic or catalyst market, the de-
mand for its supply would have a considerable impact on the co-products industry.
Column packing and support application for hydroxyapatite.
As HAP crystal structure has stable hydroxyl groups, it is an ideal packing material for
chromatographic applications. Applications include ion exchange substrate protein enrich-
ment-recovery by column chromatography.
Glues
They are made by boiling down the hooves of hooved animals such as horses, deer and
donkeys. The process is to break the hooves into small chunks and then boil the chunks in
water until all the hoof material has been liquefied. An acid is then added to create a thick
gel. The resultant product is then cooled and allowed to harden. Hoof glues have been used
for things like stiffening bow strings, stiffening and adhering fabric to wood, creating thin
lacquers to protect valuable objects as well as sealing glass into frames and sealing ceramic
containers. It is still used today in woodworking, specifically cabinetry.
Bone China
Cattle bones, transformed into fine "Bone China", have forayed into the finest dinner
services and tea sets imaginable. Bone China is a type of porcelain that is composed of bone
ash, kaolin and feldspathic material. 209 Thomas Frye first developed Bone China at his fac-
tory near Bow (London) in 1748. His factory was located very close to the cattle markets and
slaughterhouses of Essex, and hence he had easy access to animal bones. Frye used up to 45%
bone ash in his formulation. Others have said that Bone China was invented by the by the
Chinese in the seventh and eighth centuries B. C. and improved by Frye. Still others 210 believe
that the development of Bone China should be credited to Josiah Spode, who introduced it in
1797. Soon afterwards Bone China formulas were copied by factories in Liverpool, England.
Nowadays Bone China contains 35-45% bone ash in the clay formula, making the
major component a renewable resource.
4.4 Brains and spinal cords
For table use
The brain is usually prepared direct for the table rather than processed for industrial use.
Because of its soft texture, brain is blanched to firm the tissue before cooking. The mem-
branes (the arachnoid meninges and pia mater) are peeled from the brain before cooking.
In addition, brains are a source of cholesterol that is the raw material for the synthesis of
209. "Production of Bone Ash for the Manufacture of Bone China", Industrial Ceramics 1989, 843,
p . 767-770.
210. Karwatka D. (2009), "Josiah Spode and His World-Famous Pottery", techdirections, 68 (9): 12;
Badenhorst A. (2010), "Porcelain", Ceramics Technical, 3: 34.
62
Animal by-products (ABPs): origins, uses, and European regulations
vitamin D 3 and can be used as emulsifier in cosmetics. Other materials can be isolated from
the hypothalamus of the brain.
Clathrin
Cow brains could possibly be useful in creating batteries, electrodes and solar cells. Clath-
rin is present in every cell in the human body, and cows possess a vast wealth of it in their
bovine brains, making them an ideal source for the stuff. 211 Clathrin molecules are capable of
forming tiny cubes, spheres, and other shapes in test tubes. 212 Inorganic materials can like gold
and titanium dioxide can be added to the clathrin molecules. 213 Some of these materials have
been found to be able to absorb sunlight, or to split water into its components.
Melatonin
The hormone melatonin, extracted from the pineal gland, is being evaluated for the
treatment of schizophrenia, insomnia, and other problems 214 including mental retardation.
Fukui, Takagi and others 215 have isolated a novel analgesic peptide from the bovine brain.
211. To know more about see: Lindner R., E. Ungewick (1992), "Clathrin-associated Proteins of
Bovine Brain Coated Vesicles. An analysis of their number and assembly-promoting activity", The
Journal of Biological Chemistry, 267 (23): 16567-16573; Able S., E..Ungewickell (1990), "Auxilin, a
newly identified clathrin-associated protein in coated vesicles from bovine brain", The Journal of Cell
Biology, 111 (1): 19-29.
212. Heilshorn S., N. Melosh, S. Doniach, A. Spakowitz (201 1), Protein Biotemplates for Self Assembly
of Nanostructures, in Proceedings of the 2011 Biomolecular Materials Principal Investigators' Meeting
held on October 23-26, 2011 at the Westin Annapolis Hotel in Annapolis, (MD-USA), pp. 26-29;
VanDersarl J., S. Mehraeen, A. Schoen, S. Heilshorn, A. Spakowitz, N. Melosh (201 1), Mechanics of
2-D Clathrin Assembly, Proceedings of the 201 1 Biomolecular Materials Principal Investigators' Meeting
held on October 23-26, 201 1 at the Westin Annapolis Hotel in Annapolis, (MD-USA), p. 243.
213. Arora H„ C. Doty, Ye Yuan, J. Boyle, K. Petras, B. Rabatic, T. Paunesku, G. Woloschak (201 1),
Titanium Dioxide Nanocomposites, in Nanomaterials for the life Sciences, Vol.8: Nanocomposites, Edited
by Challa S. S. R. Kumar, Wiley- VCH Verlag GmbH & Co. KGaA, Weinheim; Heilshorn S. C.
(2010), Self Assembling Materials for Energy Storage and Transport, 55th Annual Report on Research
2010, Reports: DNI10.
214. People use melatonin to adjust the body's internal clock, i. e. for jet lag, for adjusting sleep-wake
cycles (shift-work disorder), and for helping blind people establish a day and night cycle. It is also used
for the insomnia; delayed sleep phase syndrome; insomnia associated with attention deficit-hyper-
activity disorder; insomnia due to certain high blood pressure medications called beta-blockers; and
sleep problems in children with developmental disorders including autism, cerebral palsy, and mental
retardation. Some people use melatonin for Alzheimer's disease, ringing in the ears, depression, chronic
fatigue syndrome, fibromyalgia, migraine and other headaches, irritable bowel syndrome, osteoporosis,
a movement disorder called tardive dyskinesia, epilepsy, as an anti-aging agent, for menopause, and for
birth control. Other uses include breast cancer, brain cancer, lung cancer, prostate cancer, head cancer,
neck cancer, and gastrointestinal cancer.
215. Fukui K„ H. Shiomi, H. Takagi, K. Hayashi, Y. Kiso, K. Kitagawa (1983), "Isolation from bovine
brain of a novel analgesic peptapeptide, neo-kyotorphin, containing the Tyr-Arg (kyotorphin) unit",
Neuropharmacology, 22 (2): 191-6; Takagi H., H. Shiomi, K. Fukui, K. Hayashi, Y. Kiso, K. Kitagawa
(1982), "Isolation of a novel analgesic pentapeptide, neo-kyotorphin, from bovine brain", Life Sciences,
31 (16-17): 1733-1736.
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Spinal cord: factor B
As we know, the spinal cord is a long, tubular bundle of nerve tissue and support cells
that extends from the brain lengthwise along the back within the vertebral canal. Pairs of
spinal nerves, originating in the spinal cord, carry impulses to and from the brain. Spinal
cord-derived growth factor-B is a member of the platelet-derived growth factor family.
Cholesterol
Cholesterol is a significant lipid in animal fats and the most abundant steroid. It can
be extracted from the spinal cord of animals and used in production of vitamin D and sex
hormones. These compounds have multiple functions in biochemical systems acting as lipids
(e.g. cholesterol), surfactants (the bile acids), and hormones etc.
4.5 Fats and fatty acids
Animal fats are made mainly from triglycerides. The product from rendering constitute
about 25% of the lipid resources used in the world. The main use of animal fats and oils are
for human and animal nutrition; the fats and oil are a dietary source of energy.
The unsaturated fatty acids (linoleic, linolenic, oleic and arachidonic acid) are nutri-
tionally important as they are necessary constituents of cell walls, mitochondria and other
intensively active metabolic sites of the living organism.
In recent years it has been suggested that a high ratio of unsaturated to saturated fatty
acids in the diet is desirable as this may lower the individual's susceptibility to cardiovascular
diseases. There is indicating evidence that a diet relatively high in saturated fats (such as those
of found in red meat) raises the level of cholesterol in the blood.
The fat content of animal carcasses vary from about 8 to 20% (the latter only in pigs).
The fatty acid composition of the fatty tissues is very different in various locations of the
body. For example, external fat ("body fat") contains a higher concentration of unsaturated
fat and is much softer than the internal fat surrounding organs, which has a high saturated
fat content. In pigs, the subcutaneous 216 fats are separated from other tissues and are suitable
for feeding, industrial uses and for meat processing (backfat, jowl fat and belly). The kidney
fat and leaf fat of pigs are used for lard production, as they are not recommended as an in-
gredient for processed meat products because of their hardness and taint.
Beef fats (tallow) are considered less suitable for further processing than pig fats due to
their firm texture and intense flavour. If used for processing, preference is given to brisket fat
and fats from young animals. Such fats are used for specific processed beef products.
Some tropical cattle breeds have a large subcutaneous fat depot, known as "hump", in
the shoulder region, that is often cut into slices and roasted as a delicacy, and sometimes used
for processed products. Buffalo fat has a white colour and is well suited for processing. The
limiting factor for utilization of tropical beef and buffalo fat is scarce availability.
Almost always mutton fat of adult animals is absolutely unsuitable for consumption due
to its intensive unpleasant flavour and taste. Fats from lamb are relatively neutral in taste and
commonly eaten with meat.
Chicken fat has a neutral taste and is suitable as a fat component for pure chicken prod-
216. Subcutaneous and intermuscular fats are known as "body fats", while another category are the
"depot-fats", located in the animal body around internal organs.
64
Animal by-products (ABPs): origins, uses, and European regulations
ucts. Chicken fat adheres to chicken muscle as intermuscular fat. The majority of chicken fat
derives from chicken skin with its high subcutaneous fat content. At present these fats are
underutilized render materials. They do not appear in the end use market.
The use of the animal fats in industrial applications, in Europe and in USA, on average,
regards the following areas with the utilization percentage:
- plastics and plasticizers 28%
- adhesives 20%
-surfactants 15%
- solvents 15%
- industrial chemicals 12%
- lubricants 8%
- agrochemicals 2%
Industrial uses for fats and fatty acids are the following:
Antifreeze, biodegradable detergents, biodiesel, bone char to filter and decolorize sugar
solutions, bone china, ceramics, chemicals, cosmetics, crayons, creams and lotions, dish and
hand soap, explosives, floor wax, fuel, hair conditioner, herbicides, industrial oil and lu-
bricants, insecticides, makeup, medicines, mink oil, oleo margarine & shortening, paints,
paraffin, rubber products, saddle soap, shaving cream, solvents, tallow for tanning.
For production of energy
Historically, tallow has had much wider energy and industrial application than protein-
based meals. The rendering process is not a new industry; the production of tallow for candles
and soap has occurred for centuries. Soap making has made major use of tallow. In the nine-
teenth century the Industrial Revolution transformed the agriculture sector. The development
of intensive livestock production led to a burgeoning disposal problem. Rendering became an
attractive solution. Early twentieth century processes, called tankage, separated the fat and water
from the protein, which was then used as a fertilizer. It was only during the the first years of the
1900s that the conversion of ABPs to meal and bone meal for animal feed became important.
The onset of World War I and II saw significant demand for rendered glycerin for the
production of explosives, specifically tri-nitroglycerin (TNT). The production of meal and
bone meal and tallow continued to increase after the war. According to the UK Department
for Environment, Food and Rural Affairs, 217 the production and use of meal and bone meal
steadily increased throughout the first half of the century. Tallow, however, was the primary
product of rendering.
Tallow can be used directly as a boiler fuel or to manufacture biodiesel. There are physi-
cal and chemical methods for transforming this product into a commercial fuel. Some sys-
tems may require filtration for fats and greases before use as boiler fuel. The tallow produced
by mortality rendering can be used as an alternative burner fuel. With good results, a mix-
ture of chicken fat and beef tallow has been blended with No. 2 fuel oil in a ratio of 33%
chicken fat/beef tallow and 77% No. 2 fuel oil. But the best way to get energy from tallow
or fats, normally, is to turn them into biodiesel.
217. UKDEFRA (2000), The BSE inquiry report, Vol. 13: Industry processes and controls, Ch. 6 render-
ing, Annex B manufacturing processes of rendering (See: http://www.bseinquiry.gov.uk/report/vol-
ume 13/ chapterj .htm) .
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Raffaella Leoci
Despite precise written sources, the concept of using vegetal oil as an engine fuel likely
dates back to Rudolf Diesel (1858-1913) who developed the first engine to run on peanut
oil, which was demonstrated at the World Exhibition in Paris in 1900. Diesel believed 218
biomass fuel to be a viable alternative to the resource consuming steam engine. Vegetable
oils were used in diesel engines until the 1920s when an alteration was made to the engine
in order to use a fraction of petroleum. That petroleum is now known as diesel fuel (in USA
diesel No. 2). In recent years, instead of diesel fuel, the Biodiesel produced from renewable
sources such as vegetable oils, animal fats such as tallow, 219 and recycled cooking oils can be
used in the engines. 220
Biodiesel, defined as the mono-alkyl esters of vegetable oils or animal fats, is produced
by transesterification with an alcohol in the presence of a catalyst. It is biodegradable and
non-toxic, and when burned has significantly fewer emissions than petroleum-based diesel.
Biodiesel functions in current diesel engines, and is a possible candidate to replace fossil fuels
as a significant supplier to the world's transport energy.
Glycerol is a major by-product of the production of that reaction. The greatest attention
at this time is the production of biodiesel fuel in the form of methyl esters. The process is
typically catalyzed by sodium hydroxide (NaOH) or potassium hydroxide (KOH) to in-
crease reaction rates. Biodiesel has a gross calorific value of about 33.3 MJ/1 and a density
of 0.88 kg/1.
A higher free fatty acid (FFA) composition of fat is likely to require more pretreatment
before biodiesel production and will generate a lower quality glycerin byproduct. Higher
levels of free fatty acids (FFA) generally mean lower quality and value of tallow. Commer-
cial operations do exist that convert FFA to biodiesel in the presence of acid-based catalysts
where the FFA content is less than 20 percent.
.Biodiesel is already in wide use around the world because it offers many advantages: it
is in liquid form and is therefore easily stored and transported; it can be blended with diesel
fuel in the same way that ethanol is blended with petrol. 221
However, biodiesel produced from tallow presents one disadvantage: its cold flow prop-
erties. Crystallization in tallow esters (biodiesel) occurs due to the high melting points of
the saturated fatty acid esters present in the biodiesel. 222 Neat (100%) methyl tallowate bio-
diesels have been shown to crystallize at significantly higher temperatures (melting point of
methyl stearate is 39.1°C) than regular diesel (i.e., up to 15°C).
218. "The use of vegetable oils for engine fuels may seem insignificant today. But such oils may become
in the course of time as important as the petroleum and coal tar products of the present time", Rudolf
Diesel, 1912 (See: Nitske W.R., C. M. Wilson (1965), Rudolf Diesel Pioneer of the Age of Power, Uni-
versity of Oklahoma Press, Norman, (OK - USA).
219. The fats produced by the rendering process can be divided into two groups: edible and inedible.
Edible fats are likely to attract a higher price in the food market. Inedible rendering products attract a
lower price and may be more suitable for biodiesel production.
220. Felizardoa P., M. J. Neiva Correiaa, I. Raposob, J. F. Mendesc, R. Berkemeierd, J. Moura Bordado
(2006), "Production of biodiesel from waste frying oils", Waste Management, 26 (5): 487-494.
221. The standard blend is 20 percent biodiesel, 80 percent diesel fuel (Paisley M. A. (2001), Biomass
Energy, in Kirk-Othmer Encyclopaedia of Chemical Technology, by John Wiley & Sons, Inc., Weinheim).
222. Papadopoulos E., S. Clarke (2005), Modification of Tallow for Better Performance as Biodiesel,
Flinders University, Adelaide, Australia.
66
Animal by-products (ABPs): origins, uses, and European regulations
Several options exist for the improvement of cold flow characteristics, 223 including
blending with regular diesel, use of branched chain alcohols, 224 and the use of additives. 225
With regard to the economic aspects, the main benefit would be the conversion of
low value, inedible rendered products to a higher value medium energy content fuel. The
estimated operating costs of biodiesel production 226 indicate that most of the operating cost
associated with typical biodiesel production is the cost of the raw material (oil/fat). The cost
of methanol, labor, catalyst, and auxiliaries was deemed to be very low. The raw material cost
was estimated as 85.8 percent of the total yearly operating costs.
Production of Hydrogen
Hydrogen is a clean fuel and feedstock to the energy and industrial chemicals indus-
tries. 227 An interesting process produces hydrogen, the aqueous-phase reforming of glycer-
ol. 228 In their study, Liu and others focused on ethylene glycol (EG), a chemical found in the
cooling system of most vehicles, and on glycerol, a by-product of the hydrolysis of fats and
oils to make biodiesel. Both EG and glycerol could be reformed to hydrogen and carbon
dioxide at 240 °C. Hydrogen yields from reforming EG and glycerol were 82% and 70%,
respectively. The benefit of this operation is the conversion of glycerol to a more valuable
product, hydrogen that can be used as a fuel in fuel cells. 229
223. Knothe G. (2008), "«Designer» Biodiesel: Optimizing Fatty Ester Composition to Improve Fuel
Properties", Energy & Fuels, 22: 1358-1364; Knothe G. (2005), "Dependence of biodiesel fuel properties
on the structure of fatty acid alkyl esters", Fuel Processing Technology, 86: 1059—1070.
224. Wang P. S., M. E. Tat, J. Van Gerpen (2005), "The production of fatty acid isopropyl esters and
their use as a diesel engine fuel", Journal of the American Oil Chemists' Society, 82 (1 1): 845-849; Miller
S. J. (201 1), US Pat. 201 10239529, Biodiesels useful for improving cloud point (06-Oct-201 1); Lee I.,
L. A. Johnson, E. G. Hammond (1995), "Use of branched-chain esters to reduce the crystallization
temperature of biodiesel", Journal of the American Oil Chemists' Society, 72 (10): 1155—1160.
225. Ryan C. C. (2008), "Additive lowers tallow-based biodiesel cloud point", Biodiesel Magazine,
October 14, 2008 (www.biodieselmagazine.com).
226. Nelson R. G., S.A. Howell, J. Weber (1994), Potential Feedstock Supply and Costs for Biodiesel Pro-
duction, presented at the sixth national bioenergy conference in Nevada, October 2-8, (www.biodiesel.
org/ resources/ reportsdatabase/reports/gen/1994l006_gen-290).
227. Gupta R. B. (2009), Hydrogen Fuel: Production, Fransport, and Storage, CRC Press, Florida (USA).
228. Liu B., Y. Zhang, J. W. Tierney, I. Wender (2005), Hydrogen by Catalytic Reforming of Glycols, in
"CI Chemistry for the Production of Ultra-Clean Liquid Transportation Fuels and Hydrogen", Semi-
annual six-month report. Research conducted October 1, 2004-March 31, Prepared by the Consor-
tium for Fossil Fuel Science, Gerald P. Huffman, Director CFFS/University of Kentucky, Lexington,
KY, pp. 75-77.
229. Fuel cells are electrochemical devices that convert chemical energy directly to electricity. They of-
fer a significant inherent advantage over typical internal combustion engine where efficiency is lost due
to the conversion of stored chemical energy first to heat energy, then to mechanical energy, and finally
to electricity. Fuel cells are not subject to Rankine/Carnot cycle efficiency limitations and are energy
carriers, like batteries. In theory, a fuel cell can continue to produce power indefinitely if a fuel stream
such as hydrogen is constantly provided.
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4.6 Glands and organs
Animal glands are traditionally used as medicine in many Western and Eastern coun-
tries, including China, India, South Korea, New Zealand and Japan.
The more exploited are liver, pituitary, thyroid, pancreas, stomach, adrenal glands, kid-
ney, spleen, lungs, thymus, thyroid, and ovary. Different animals have different glands that
are important. Their function also depends on the species, sex and age of the animal. They
are collected only from healthy animals.
a) Adrenal glands
Mammals adrenal glands are endocrine glands that sit at the top of the kidneys. They
regulate the response to stress by secreting hormones and are divided into two parts: the in-
ner portion (medulla) which secretes at least 20 steroids that are essential for maintaining
life (among these it secretes are epinephrine and norepinephrine); and the outer portion (the
cortex) which manufactures the hormones cortisone and aldosterone.
Adrenal extract is a chemical made from the inner portion of the adrenal glands of
slaughtered cows, pigs, and sheep. It is a trisubstituted benzene and is used as a heart stimu-
lant. By mouth, adrenal extract is used for low adrenal function, fatigue, asthma, stress, severe
allergies, lowered resistance to illness, certain skin conditions such as eczema and psoriasis,
and rheumatoid arthritis. Recent findings 230 show its role in the treatment of anaphylaxis, 231
croup, 232 bronchiolitis, 233 and as an adjunct to local anesthesia. 234
Steriods from the cortex of the adrenal gland regulate the body's utilization of nutrients
such as fat, carbohydrates, water and minerals. Steriods extracted from cattle, pigs or sheep
are used as anti-inflammatory agents and for the treatment of shock and asthma. One such
steroid, corticosterone, derived from bovine adrenals, is extracted directly from the cortex of
the adrenal glands.
Adrenalin, without the "e", was originally used as a trademark for a product made by an
American pharmaceutical firm Parke, Davis & C 235 . The usage of the word, with or without
the "e", however, was not very consistent; thus the term's use seems interchangeable.
Epinephrine and nor-epinephrine, extracted from the medulla of cattle, pigs, and sheep,
are glande extracts used to stop hemorrhaging, stimulate heart action and overcome shock.
230. See also: Walker D. M. (2009), Update on epinephrine (adrenaline) for pediatric emergencies, Lip-
pincott Williams & Wilkins, Inc., Wolters Kluwer N.V.
231. Garvey L. H., B. Belhage, M. Kroigaard, B. Husum, Hans-Jorgen Mailing, H. Mosbech (201 1),
"Treatment with epinephrine (adrenaline) in suspected anaphylaxis during anesthesia in Denmark",
Anesthesiology, 115(1): 11-6.
232. Racemic epinephrine has historically been used for the treatment of croup. See: Thomas L. P.,
L. R. Friedland (1998), "The cost-effective use of nebulized racemic adrenaline in the treatment of
croup", American journal of Emergency Medicine, 16 (1): 87-89.
233. Menon K., T. Sutcliffe, T. P. Klassen (1995), "A randomized trial comparing the efficacy of epi-
nephrine with salbutamol in the treatment of acute bronchiolitis", The Journal of Pediatrics, 126(6):
1004-1007.
234. Wong J. K. (2001), "Adjuncts to Local Anesthesia: Separating Fact from Fiction", Journal of the
Canadian Dental Association, 67: 391-7.
235. The Parke-Davis Research Laboratory is an American National Historic Landmark; the surrounding
Parke-Davis and Company Pharmaceutical Company Plant is on the National Register of Historic Plac-
es. Parke-Davis was acquired by Warner-Lambert in 1970, which in turn was bought by Pfizer in 2000.
68
Animal by-products (ABPs): origins, uses, and European regulations
The molecular formula for epinephrine is C 9 H [3 N0 3 . Epinephrine is derived from ty-
rosine, an aminoacid. Epinephrine is sometimes referred to as a catecholamine as it contains
the catechol moiety. This is a part of the molecule that contains the group C 6 H 4 (OH) 2 .
b) Kidney
Animal kidneys (from catties, pigs, or sheeps) after having been removed from the fatty
capsule which holds them in place, may be cooked whole or in slices. Kidney is generally
broiled, grilled, or braised.
For transplantation
Kidney was one of the first organs to be used for transplantation. Historically, kidney
transplantation is thought to have originated early in the twentieth century with several at-
tempts of xenografting and experimental works on vascular sutures. 236
The first successful experimental kidney transplant was performed at the Vienna Medi-
cal School in Austria in 1902. The surgeon Ullmann 237 reported the first case of renal auto-
transplantation performed in the neck of a dog. In 1906, M. Jaboulay, 238 in Lyon, used
kidneys from pigs and goats to attempt xenotransplantation in a human patient. According
to records the first kidney transplant experiments were performed in France in 1909 on hu-
mans using animal kidneys. 239 In this experiment a surgeon inserted slices of rabbit kidney
into a child suffering from kidney failure. Although "the immediate results were excellent"
the child died about 2 weeks later. In 1909, Ernst Unger used en-bloc Macaccus kidneys in
humans that rapidly failed, due to an unknown hyperacute vascular rejection. 240 Scientists
of the time believed kidney xenotransplants were possible, but their success was limited by
unknown "biochemical barriers," which prevented long-term kidney survival.
The site of the first successful human-to-human kidney transplant was at the Peter Bent
Brigham Hospital in Boston, Massachusetts, where, in 1954, Joseph E. Murray transplanted
236. Kiiss R., P. Bourget (1992), Une histoire illustree de la greffe d'organes. La grande aventure du
siecle, Laboratoires Sandoz, Rueil-Malmaison (France); Taniguchi S., D. K. Cooper (1997), "Clinical
xenotransplantation: past, present and future", Annals of The Royal College of Surgeons of England, 79
(1): 13-19; Cooper D. K. C. (2012), "A brief history of cross-species organ transplantation", Baylor
University Medical Center Proceedings, 25 (1): 49-57.
237. Emerich Ullmann was Chief Surgeon at Spital der Baumherigen Schwetern, Germany.
238. Schlich T. (201 1), "The origins of organ transplantation", The Lancet, 378 (9800): 1372 - 1373.
239. Princeteau M. J. (1905), "GrefFe renale", Journal de Medicine de Bordeaux, 26: 549; Reemlsma
K. (1969), "Renal heterotransplantation from nonhuman primates to man", Annals of the New York
Academy of Sciences, 162: 412-8; Deschamps J. -Yves, F. A. Roux, P. Sa'i, E. Gouin (2005), "History of
xenotransplantation", Xenotransplantation, 12 (2): 91-109.
240. For more notices see: Nagy J. (1999), "A note on the early history of renal transplantation:
Emerich (Imre) Ullmann", American Journal of Nephrology, 19 (2): 346-9; Jorge O. (201 1), A. Jason
(Edtrs.), Understanding the Complexities of Kidney Transplantation, InTech, Rijeka, Croatia (available at
www.intechopen.com); Brent L. (1997), A history of transplantation immunology, Academic Press, San
Diego, (CA-USA); Doyle A. M., R. I. Lechler, L. A. Turka (2004), "Organ Transplantation: Halfway
through the First Century" , Journal of the American Society of Nephrology, 15 (12): 2965-2971; Schlich
T. (2010), The origins of organ transplantation: surgery and laboratory science, 1880s — 1930s, University
of Rochester Press, Rochester, (NY-USA).
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Raffaella Leoci
a kidney that was donated by the patient's twin. 241 Many other efforts followed but all failed
with short survival of the affected, 242 except one. The only success was the nine-month sur-
vival of a chimpanzee kidney transplanted into a human by Reemtsma and colleagues. 243 No
one knows why this survival occurred, in the 1960s there were no powerful immunosuppres-
sive agents, and no known immunological or genetic manipulations of donor or recipient.
Xenotransplantation was seen by some as an opportunity and by others as a danger. It
could help overcome the shortage of organs from human donors, but it raised many ques-
tions about safety, ethics and human nature. 244 As a consequence many researchers settled on
the use of pigs as potential source animals for xenotransplantation. 245 In most recent years,
researchers are experimenting with producing organs from genetically modified animals.
With the advent of genetic engineering and cloning technologies, pigs are currently available
with a number of different manipulations that protect their tissues from the human immune
response. The use of genetic engineering has resulted in significant improvement in survival
time for a pig organ in a non-human primate receiving immunosuppression. 246
241. Morris P. (2013), "Joseph E. Murray (1919-2012)", Nature, 493 (7431): 164; Guild W. R„ J. H.
Harrison, J. P. Merrill, J. Murray (1955), "Successful homotransplantation of the kidney in an identi-
cal twin", Transactions of the American Clinical and Climatological Association, 67: 167—173.
242. Mohacsi P. J., J. F. Thompson, S. Quine (1998), "Attitudes to xenotransplantation: scientific
enthusiasm, assumptions and evidence", Annals of Transplantation, 3 (2): 38-45.
243. Reemtsma K., B. H. McCracken, J. U. Schlegel, M. A. Pearl, C. W. Pearce, C. V. DeWitt, P. E.
Smith, R. L. Hewitt, R. L. Flinner, O. Jr Creech (1964), "Renal hetero xenotransplantation in man",
Annals of Surgery, 160: 384—410.
244. Daar A. S. (1999), "Animal-to-human organ transplants - a solution or a new problem?", Bulletin
of the World Health Organization, 77 (1): 54-61.
245. Cooper D. K. C, Y. Ye, J. L. L. Rolf, N. Zuhdi (1991), The Pig as Potential Organ Donor for
Man, in "Xenotransplantation. The Transplantation of Organs and Tissues Between Species", D. K. C.
Cooper, E. Kemp, K. Reemtsma, D. J. G. White (Edts.), 1st ed., Springer- Verlag, Berlin, pp. 481-500;
Hammer C, R. Linke, E Wagner, M. Diefenbeck (1998), "Organs from animals for man", Interna-
tional Archives of Allergy and Immunology, 116: 5-21.
246. Cozzi E., D. J. G. White (1995), "The generation of transgenic pigs as potential organ donors
for humans", Nature Medicine, 1: 964 - 966; Lin S. S., B. C. Weidner, G. W. Byrne, L. E. Diamond,
J. H. Lawson, C. W. Hoopes, L. J. Daniels, C. W. Daggett, W. Parker, R. C. Harland, R. D. Davis,
R. R. Bollinger, J. S. Logan, J. L. Piatt (1998), "The role of antibodies in acute vascular rejection of
pig-to-primate cardiac transplants", The Journal of Clinical Investigation, 101 (8): 1745-1756; Cozzi
E., F. Bhatti, J. R. Bradley, G. Chavez, P. J. Friend, M. Goddard, D. Ostlie, M. Schmoeckel, K. G. C.
Smith, S. Thiru, C. Vial, J. Wallwork, D. J. G. White, A. Zaidi (2000), "Long-term survival of non-
human primates receiving life-supporting transgenic porcine kidney xenografts", Transplantation, 70
(1): 15-21; Vial C. M., D. J. Ostlie, F. N. Bhatti, E. Cozzi, M. Goddard, G. P. Chavez, J. Wallwork,
D. J. White, J. J. Dunning (2000), "Life supporting function for over one month of a transgenic
porcine heart in a baboon", The Journal of Heart and Lung Transplantion, 19 (2): 224-229; Loss M.,
B. Vangerow, J. Schmidtko, R. Kunz, A. Jalali, H. Arends, M. Przemeck, H. Riickholt, M. Leuwer, F.
J. Kaup, S. Rensing, E. Cozzi, D. J. White, J. Klempnauer, M. Winkler (2000), "Acute vascular rejec-
tion is associated with systemic complement activation in a pig-to-primate kidney xenograft model",
Xenotransplantation, 7 (3): 186-96; Bhatti F. N., M. Schmoeckel, A. Zaidi, E. Cozzi, G. Chavez, M.
Goddard, J. J. Dunning, J. Wallwork, D. J. White (1999), "Three-month survival of HDAF transgenic
pig hearts transplanted into primates", Transplantation Proceedings, 31 (1-2): 958-66; Diamond L. E.,
C. M. Quinn, M. J. Martin, J. Lawson, J. L. Piatt, J. S. Logan (2001), "A human CD46 transgenic
pig model system for the study of discordant xenotransplantation", Transplantation, 71 (1): 132-42;
70
Animal by-products (ABPs): origins, uses, and European regulations
The first batch of genetically modified pigs was born in the second half of 20 1 1 . Genetic
engineering experts at the Nanjing Medical University (in eastern China's Nanjing) recently
announced that these pigs are expected to provide suitable organs for transplant into human
bodies and to ease the nation's shortage of transplant organs. Researchers at that Medical
University said that pig-to-human heterografts are expected to undergo clinical trials within
two to three years and that the exact time needed depends on the type of organs concerned.
c) Liver
The liver is the largest gland in animals. Liver from lamb, veal calves, pigs and young
cattle are the most widely used edible organ. Collected only from healthy animals, liver is
used in many processed meats, such as liver sausage and liver paste, especially in the United
States and Europe. In theses countries consumers generally prefer livers from pigs because it
has a lighter flavor and texture.
Liver extract
The liver of mature cattle weighs about 5 kg, while that of a pig weighs approximately
1.4 kg. 247 For use as raw material in the pharmaceutical industry, liver extract 248 is produced
by mixing raw ground liver from pigs and cattle with slightly acidified hot water. The stock is
concentrated in a vacuum to make a paste. 249 That paste has longevity as a source of vitamin
B 12 , as a nutritional supplement used to treat various types of anemia, for production of red
blood cells, as a homeopathic medicine, and as a micro nutrient.
Heparin can be extracted from the liver and used as an anticoagulant to prolong the
clotting time of blood, to thin the blood, and to prevent blood clotting during surgery and
in organ transplants.
For transplantation
The first successful human liver transplant was performed by Starzl and colleagues at
the the University of Colorado in 1967. The recipient lived for about a year before dying
from a recurrence of liver cancer. In the 1990s, Starzl and colleagues 250 transplanted baboon
livers into one patient. This patient survived for twenty-six days. The operation marks the
first known baboon-to-human liver transplant in the world. The baboon was selected as a
donor because it is not an endangered species, and can be bred safely and easily in captiv-
ity. A baboon pathogen (cytomegalovirus) was apparently transferred to the patient, even
Chang A. T., J. L. Piatt (2009), "The role of antibodies in transplantation", Transplantation Reviews,
23 (4): 191-198.
247. To know more about see: Boxenbaum H. (1980), "Interspecies variation in liver weight, hepatic
blood flow, and antipyrine intrinsic clearance: Extrapolation of data to benzodiazepines and phenyto-
in" ', Journal of Pharmacokinetics and Biopharmaceutics, 8 (2): 165-176.
248. Liver extract is a dietary supplement made from animal liver, often pig or cow, and is commonly
sold either as a freeze-dried powder or a concentrated liquid. It contains vitamin B ]2 , folic acid and
iron.
249. Fenger E, Production of liver extract, United States Patent 2045266.
250. Starzl T. E., J. J. Fung, A. G. Tzakis, S. Todo, A. J. Demetris, I. R. Marino, H. Doyle, A. Zeevi, V.
Warty, M. Michaels, S. Kusne, W. A. Rudert, M. Trucco (1993), "Baboon-to-human liver transplanta-
tion", The Lancet, 341 (8837): 65-71.
71
Raffaella Leoci
though this did not result in a disease process. 251 However, in the patient there was evidence
of an adequately functioning liver mass, sufficient to sustain life. The baboon liver led to the
presence of baboon proteins synthesized by the liver; in some cases those proteins assumed
the blood levels characteristic of the baboon and not of the human.
On June 28, 1992, Starzl complete an 11-hour transplant operation of a baboon liver
into a 35-year-old male, but the patient died after 71 days as doctors tried to wean him from
a ventilator. From 1966 to 1973 Tom Starzl transplanted three livers from chimpanzees to
children. All died within two weeks. 252 After the last transplant, protesters picketed Starlz's
house calling him "Tom FrankenStarzl".
In the meantime, scientists continue to investigate pig's liver. In 1993 Leonard Ma-
kowka put a pig liver into a human and failed. 253 Currently, future liver xenotransplantation
hopes rest on animals, particularly genetically modified pigs. Pigs could provide an alterna-
tive source of tissue and cells, but the immunological challenges and other barriers associated
with xenotransplantation need to be overcome. Transplantation of organs from genetically
modified pigs into non-human primates is now not substantially limited by hyperacute, or
acute antibody-mediated, or cellular rejection; other issues have become more prominent,
such as development of thrombotic microangiopathy in the graft or systemic consumptive
coagulopathy in the recipient. 254
Genetically modified pig liver 255 is used for ex vivo perfusion in the treatment of fulmi-
nant liver failure.
d) Pancreas
Pancreatic tissue is present in all vertebrate species, and its form and arrangement vary
widely. In teleosts, and a few other species (such as rabbits), there is no discrete pancreas at
all, with pancreatic tissue being diffusely distributed across the mesentery and even within
other nearby organs, such as the liver or spleen.
Juices for metabolic use
The pancreas, as do other glands of the animal body, produces juices for metabolic
use that help break down food, and it produces hormones that help control blood sugar
levels. Juices are a source of raw material. Among these substances are insulin, glucagon,
251 . Michaels M. G., F. J. Jenkins, K. St George, M. A. Nalesnik, T. E. Starzl, C. R. Rinaldo Jr. (2001),
"Detection of infectious baboon cytomegalovirus after baboon-to-human liver xenotransplanta-
tion", The Journal of Virology, 75 (6): 2825-8.
252. After Tom Starlz' failed xeno transplants public and professional attitudes hardened saying that
humans shouldn't be used for virtual experiments until further progress was made on reducing im-
mune reaction to animal organs.
253. Cooper D. K. C, R. P. Lanza (2000), Xeno: The Promise of Transplanting Animal Organs Into
Humans, Oxford University Press, N. Y., p. 41.
254. Ekser B., M. Ezzelarab, H. Hara, D. J. van der Windt, M. Wijkstrom, R. Bottino, M. Trucco, D.
K. C. Cooper (2001), "Clinical xenotransplantation: the next medical revolution?", Transplantation,
71 (1): 132-42; Elliott, R. B. (2011), "Towards xenotransplantation of pig islets in the clinic", Curr
Opin Organ Transplant, 16 (2): 195-200..
255. Diamond L. E., C. M. Quinn, M. J. Martin, J. Lawson, J. L. Piatt, J. S. Logan (2001), "A human
CD46 transgenic pig model system for the study of discordant xenotransplantation", ibid., 132-42.
72
Animal by-products (ABPs): origins, uses, and European regulations
pancreatin, somatostatin, pancreatic polypeptide etc. Major products such as pepsin, rennin
and other digestive enzymes, lipase and trypsin enzymes extracted from the pancreas are all
medically significant products. Chymotrypsin and trypsin are used to improve healing after
surgery or injury.
Insulin
Insulin is a hormone consisting of two linked polypeptide chains, which regulates sugar
metabolism and is used in the treatment of diabetes. The structure of human insulin is
similar but not identical to the bovine and porcine insulin produced by the Meat Industry.
The attempts to produce insulin outside the human body, dates back to 1921 when
Banting and Best, 256 and Macleod, 257 successfully purified insulin from a dog's pancreas.
Though insulin has been referenced as one of the prime pharmaceutical products de-
rived from ABPs, it is now synthesized by other procedures. 258 This is true for a number
of other pharmaceuticals, but reliance on the natural production and extraction is still an
important source of medical treatment and prevention compounds.
Pig insulin has been used to treat diabetes in humans until the 1980s, making the pigs
potentially good islet donors for humans with the disease. Insulin from pigs was routinely
given to patients with diabetes, until DNA technology enabled pharmaceutical companies
to manufacture human insulin. In a recent study, 259 the researchers transplanted clusters of
embryonic pig pancreatic cells into 1 0 diabetic rats that could not produce insulin on their
own and that had very high glucose levels. The cells were retrieved from the pig embryos
early in their development, which was believed to make them both "invisible" to the rats'
immune system and also not to induce a state of immune tolerance. Those who receive hu-
man islet transplants must take anti-rejection drugs for the rest of their lives, so essentially
they are trading daily insulin shots for immune-suppression drugs, which carry their own
risks. Pig cells could overcome the shortage of human islets available from deceased donors
and the need for transplant patients to take anti-rejection drugs for life.
Since about 20 years are ago, immune protective devices containing pancreatic islets
designed to treat insulin-dependent diabetes mellitus have been studied and tested for their
ability to provide glycemic control without immunosuppression. 260 The device consists of
256. Best C. H., D. A. Scott (1923), "The Preparation of Insulin", The Journal of Biological Chemistry,
57 (3): 709-723.
257. Simoni R. D., R. L. Hill, M. Vaughan (2002), "The Discovery of Insulin: the Work of Frederick
Banting and Charles Best", The Journal of Biological Chemistry, 111 (26): 31-32.
258. In 1982, the Eli Lilly Corporation successfully produced a human insulin, that became the first
approved genetically engineered pharmaceutical product. Human insulin is grown in the lab inside a
common bacteria, Escherichia coli. Though it is by far the most widely used type of bacterium, yeast
can also be used as a substitute.
259. Rogers and others, at Washington University School of Medicine, have alleviated the diabetic in
rats using transplants from both embryonic and adult pigs. The rats adopted the pig transplants as their
own and produced enough insulin to control their blood sugar, without the need for anti-rejection
drugs (See: Rogers S. A., T. Mohanakumar, H. Liapis, M. R. Hammerman (2010), "Engraftment of
cells from porcine islets of Langerhans and normalization of glucose tolerance following transplanta-
tion of pig pancreatic primordia in non-immune suppressed diabetic rats" , American Journal of Pathol-
ogy, 177 (2): 854-864).
260. Maki T, C. S. Ubhi, H. Sanchez-Farpon, S. J. Sullivan, K. Borland, T. E. Muller, B. A. Solomon,
73
Raffaella Leoci
a chamber through which passes a copolymer membrane connected to standard vascular
grafts. Islets are placed inside the chamber but are outside of the blood stream. Further
research has been conducted using rats. 261 In other studies 262 pig pancreatic islets (encapsu-
lated) have been used for the treatment of diabetic dogs. Preliminary obtained data suggests
that pig islet microencapsulation achieved metabolic control in type I diabetic dogs without
the risk of immunosuppression using one or two procedures per year.
Glucagon
Glucagon is a 29-amino acid polypeptide, 263 named in 1923, from the Greek ylvjKOi;. 264
Unlike insulin, glucagon extracted from the pancreas is used to increase blood sugar, to lower
blood sugar caused by alcoholism, and to treat insulin overdoses.
Bovine glucagon 265 has the same amino acid composition as human 266 and porcine
glucagon. Glucagon can be produced synthetically 267 but as the structures of bovine, porcine
and human glucagon are identical, that synthetic has no advantage over natural glucagon.
The process for recovering glucagon from pancreas glands has been the subject of nu-
merous patents. 268
Pancreatin
Pancreatin is a mixture of digestive enzymes consisting primarily of amylase, lipase and
protease. A mixture of those pancreatic juice enzymes, extracted from animals such as cattle
W. L. Chick, A. P. Monaco (1991), "Successful treatment of diabetes with the biohybrid artificial
pancreas in dogs", Transplantation, 51 (1): 43-51; MakiT., C. J. Mullon, B. A. Solomon, A. P. Monaco
(1995), "Novel delivery of pancreatic islet cells to treat insulin-dependent diabetes mellitus", Clinical
Pharmacokinetics, 28 (6): 471-82.
261. See for example: Rogers S. A., F. Chen, M. Talcott, M. R. Hammerman (2004), "Islet cell engraft-
ment and control of diabetes in rats after transplantation of pig pancreatic anlagen," American Journal
of Physiology, 286 (4): E502-E509.
262. See: Abalovich A. G., M. C. Bacque, D. Grana, J. Milei (2009), "Pig pancreatic islet transplanta-
tion into spontaneously diabetic dogs", Transplantation Proceedings, 41 (1): 328-30.
263. Bromer W. W., L. G. Sinn, A. Staub, O. K. Behrens (1957), "The amino acid sequence of gluca-
gon", Diabetes, 6 (3): 234-8.
264. To know more about see: Kieffer T. J., J. F. Habener (1999), "The Glucagon-Like Peptides",
Endocrine Reviews, 20 (6): 876-913.
265. Bromer W. W., M. E. Boucher, J. E. Koffenberger (1971), "Amino acid sequence of bovine glu-
cagon", Journal of Biological Chemistry, 246 (9): 2822-2827.
266. Thomsen J., K. Kristiansen, K. Brunfeldt, F. Sudky (1971), "The amino acid sequence of human
glucagon", FEES Letters, 21 (3): 315-319.
267. Lundt B. E, F. C. Groenvald, N. L. Johansen, J. Markussen (1979), "Synthesis of glucagon",
Research Disclosure, 181: 246-247.
268. Among others see: Maskalick D. G., M. T. Anderson - US Patent 4,617,376, 1986; Maskalick
D. G., M. T. Anderson - EP Patent 0,207,727, 1987; Smith M. R. - US Patent 3,715,345, 1973;
Stilz J. G., R. L. Jackson - US Patent 4,033,941, 1977; Moody A. J., L Turn, K. D. Jorgensen - US
Patent 4,405,608, 1983; Ditte Riber (Frederiksberg, DK), Eddi Meier (Vaerlose, DK), Trine Skov-
lund Ryge (Frederikssund, DK), Jens Rosengren Daugaard (Virum, DK) - Patent application number:
20100204105, 2010; Drucker D. J. (Toronto, CA), A. E. Crivici (San Diego, CA, US), M. Sumner-
Smith (Bolton, CA) - Patent application number: 20110009320, 2011.
74
Animal by-products (ABPs): origins, uses, and European regulations
or hogs is used as a digestive aid. Supplemental pancreatic enzyme preparations are provided
to patients with conditions of pancreatic exocrine deficiencies such as chronic pancreatitis
and cystic fibrosis. 269 Pancreatin is also used for improving digestion of fatty foods, to assist
specifically with the digestion of carbohydrates, lipids and proteins and it is used in pancrea-
titis control. 270
As capsules, the enzyme found on the open market might help people suffering from:
• Cystic fibrosis
• Different forms of cancer
• Heart disease
• Viruses
Pancreatic enzyme preparations, generically called pancreatin, are not alike. Rather, they
present a broad variety of pancreatin composition. 271
Pancreatin is produced by activation, extraction, precipitation, degreasing, drying, and
crushing of milled pig pancreas. Porcine pancreas is preferred for the industrial manufacture
of Pancreatin for the following reasons: it is commercially available in substantial quanti-
ties; it contains high Protease, Amylase and Lipase activity (provided the glands were frozen
immediately after slaughtering); and the composition is similar to that of human pancreas.
Some methods of preparation are covered by patents. A dry pancreatin preparation 272 is pro-
duced from a still moist pancreatin mass, which is obtained after extraction with a solvent or
solvent mixture. Before the solvents are finally removed, it is then treated,, for a short time
in a vacuum cutter until the mass has been divided and shaped into small spherical particles,
and then it is finally dried.
e) Pituitary glands
The pituitary gland (or hypophysis), known as the "master gland" of the endocrine system,
is located at the base of the brain and produces several species of hormones that help in the
control of many processes. 273 Among the most important hormones, there are the Growth-
promoting Hormone (GH) (excess of GH can lead to gigantism), the thyroid stimulating
hormone (TSH), the mammary stimulating hormone and the adrenal-cortex stimulating hor-
mone (ACTH) . Other types of hormones, secreted from the pituitary gland, are responsible for
controlling blood pressure, breast milk production, conversion of food into energy, tempera-
ture regulation, sex organ functions (in both males and females), and pain relief, etc.
Major hormonal changes emerge during pregnancy. The pituitary gland is one of the
organs most affected with altered anatomy and physiology. Due to physiological changes in
269. Ferrone M„ M. Raimondo, J. S. Scolapio (2007), "Pancreatic enzyme pharmacotherapy", Phar-
macotherapy, 27 (6): 910-20.
270. Read more at http://www.drugs.com/cdi/pancreatin.html.
271 . Lohr J. M , F. M. Hummel, K. T. Pirilis, G. Steinkamp, A. Korner, F. Henniges (2009), "Properties
of different pancreatin preparations used in pancreatic exocrine insufficiency", European Journal of Gas-
troenterology & Hepatology, 21 (9): 1024-1031.
272. Atzl G., F. Langer, H. Polleres - US Patent 5,861,177, 1999; Wilson Pharmaceutical & Chemical
Corporation - 3,956,483, 1971; Kali-Chemie Pharma GmbH - US Patent 4,019,958, 1977; CAN
Technologies, Inc. US Patent 7,153,504 - 2006.
273. Melmed S. (2011), The pituitary, Academic Press, third edition, London; Charkravarthy R. Kan-
nan (1987), The pituitary gland, Plenum Medical Book Co, New York.
75
Raffaella Leoci
the pituitary and target hormone levels, binding globulins, and placental hormones, hormo-
nal evaluation becomes more complex in pregnant women. 274
ACTH, the main hormone extracted from the pituitary, is used as a treatment for rheu-
matism, arthritis, eye inflammation and multiple myeloma.
The induction of amphibians ovulationwith the use of hormones has been used since
the early 20 th century. Some amphibians have never reproduced in captivity without hor-
mones and by using hormonal induction many species can be reproduced at will. 275 Pituitary
hormones have been very successful in encouraging ovulation in some organisms when puri-
fied hormones did not succeed. But there are disadvantages: they may not be commercially
available in all countries and have the potential to transmit disease.
Anyhow, artificially injected hormones in animals can be profitable for the farmers as
they help the animals to gain weight as well as increase the amount of milk produced. 276
Various hormones such as "Bovine Growth Hormone" (BGH) estrogen can be artificially
introduced into the cow pituitary glands to increase the production of milk. There are ques-
tions about the possible side effects on the human body after consuming the hormones in
meat and dairy products. 277 The practice of injecting hormones in cattle and poultry is very
common, and down the line can be harmful for humans and animals, as well as the environ-
ment, and might cause side effects like mastitis. 278
BGH, also called "bovine somatotropin" (BST), produced in the pituitary glands of
dairy cows, is a naturally occurring protein hormone in milk, which stimulates the liver to
produce insulin-like growth factor-I. 279 In the early stages of a calf's development, BST acts
as a growth hormone. During lactation, it serves to mobilize body fat for use as energy and
diverts feed energy more toward milk production than toward tissue synthesis. For these
reasons a large number of cattle in the U.S. and in the E. U. are supposedly being fed BGH
in order to increase milk production.
In the past GH was extracted from human pituitary glands and given to deficient chil-
dren and cattle. Soon with the increase in consumption, the quantity of GH produced
from pituitary glands was insufficient to meet the demand. As a consequence biotechnology
research received a boost to produce hormones using genetically engineered bacteria. The
274. Karaca Z., F. Tanriverdi, K. Unluhizarci, F. Kelestimur (2010), "Pregnancy and pituitary disor-
ders", European Journal of Endocrinology, 162 (3): 453-475.
275. One of the great advantages of hormonal induction is the production of oocytes without fertili-
sation by males. Several classes of compounds are used as hormones: compounds such as pimozide,
pituitary extracts, etc.
276. Today, the cattle are often raised by using artificial hormones to increase the growth rate as well as
the body mass. Artificially induced hormones in animals can increase the dairy as well as meat produc-
tion. Read more at: http://www.buzzle.com/articles/hormones-in-food.html.
277. After thorough research, the SCVPH (Sub-Committee on Veterinary Public Health) came to the
conclusion that "no acceptable daily intake could be established for any of these hormones" and those
who eat food products having these hormonal residues are at a great risk of severe hormonal imbalance
as well as various types of cancer.
278. Udder infection causing pus that can later get into the milk that we consume. As a consequence,
they are given some type of antibiotics that cause more trouble.
279. The structure of human somatotropin differs from BGH, and the latter seems not to be biologi-
cally active in humans. See: Parodi P. W. (2005), "Dairy product consumption and the risk of breast
cancer", Journal of the American College of Nutrition, 24 (6): 556S-568S.
76
Animal by-products (ABPs): origins, uses, and European regulations
genes responsible for production of BST in cattle were identified in bovine tissue cells; they
cause the pituitary cells to produce the biological product BST. These genes were isolated
and inserted into specific bacteria as part of a plasmid. As these altered bacteria replicated,
the new genes are also replicated and passed along to all new bacteria that essentially become
little "manufacturing plants" able to produce BST in large quantities. In milk the synthetic
form of BST cannot be distinguished from natural form. Somatrem and Somatropin are
man-made versions of human growth hormone. 280
GH is now produced synthetically and given to both children and adults for a variety of
reasons, 281 but GH therapy is a focus of social and ethical controversies.
f) Spleen
"The spleen has been considered a mysterious organ since classical times. For many years
after its existence became known, it appeared to have no function. Now we know this to be
far from the truth, yet many individuals survive perfectly well in the absence of a functioning
spleen, following surgery or as a result of diseases causing hyposplenism". 282
Spleen, as it is well known, is a large, highly vascular lymphoid organ present in virtu-
ally all vertebrate animals. It serves to store blood, disintegrate old blood cells, filter foreign
substances from the blood, and produce lymphocytes. It also holds a reserve of blood that
can be released into the circulatory system to meet a sudden demand, such as bleeding
(hemorrhage).
Spleen's main function is to act as a filter for the blood. It recognizes and removes old,
malformed, or damaged red blood cells. When blood flows into the spleen, the red blood
cells must pass through a labyrinth of narrow passages. Healthy blood cells can pass through
the spleen and continue to circulate throughout the bloodstream, while blood cells that are
not able to pass will be broken down in the spleen by macrophages. Useful components from
the old cells, such as iron, 283 will be saved Unfortunately no one completely understands the
spleen's functions. 284
Another function of the spleen is the accumulation of blood. Depending on the amount
accumulated, the spleen is able to get wider or narrower, to meet the needs of the bodies.
When it is expanded it can hold a great reserve of blood in order to respond to further re-
quests for blood (due to a trauma, during transfusions, etc.).
The spleen also plays an important role in the immune system, which allows you to fight
infection. When it detects faulty red blood cells, the spleen identifies and pick out enemy or
unwelcome micro-organisms (bacteria, viruses, etc.) in the blood.
280. Bramnert M., M.Segerlantz, E. Laurila, J. R. Daugaard, P. Manhem, L. Groop (2003), "Growth
hormone replacement therapy induces insulin resistance by activating the glucose-fatty acid cycle", The
Journal of Clinical Endocrinology & Metabolism, 88 (4): 1455-1463.
281. Man-made growth hormone may be used in children who have failure to grow normally. Among
the reasons there are the inability to produce enough growth hormone, kidney disease, Prader-Willi
Syndrome, and Turner's syndrome. Growth hormone is also used in adults to treat growth failure and
to treat weight loss caused by acquired immunodeficiency syndrome (AIDS).
282. Wilkins B. S. (2002), "Historical Review", 2002 Blackwell Science Ltd, British Journal of Hae-
matology, 117 (2): 265-274.
283. Spleen stores iron in the form of ferritin or bilirubin, and returns the iron to the bone marrow.
284. Wilkins B. S. (2002), "Historical Review", ibid.: 265.
77
Raffaella Leoci
The spleen continues to produce red blood cells throughout life in most vertebrates.
Many mammals possess tiny spleen-like structures known as haemal nodes throughout the
body, which, it is presumed, have the same function as the spleen proper.
In addition to food
Spleen seems also to be a good food. For instance, spleen is minced and used in pies and
as a flavoring in the United Kingdom, and as an ingredient in processed meat in the United
States. To be safely eaten it must, of course, be perfectly healthy. 285 But everyone knows that
it is subject to various diseases. One thinks of hog cholera, tuberculosis, charbon, and other
diseases of the same group. If it is not perfectly fresh, the large number of blood-corpuscles in
the spleen pulp renders it peculiarly prone to decomposition. When cooked, it keeps better.
Its softness and the necessity of eating spleen only when it is quite fresh are reasons why
it is unmarketable in their natural state and can only be obtained at the slaughter-houses.
Ferritin
In addition to food, spleen can be used to obtain ferritin from horse (equine) and cattle
(bovine) that are used in the treatment of iron-deficiency anemia. "Spleen extract" is given
"as replacement therapy" in cases where the spleen has been surgically removed and or isn't
working properly.
For stimulating the immune system
Spleen extract is also used for treating "autoimmune" diseases such as celiac disease,
systemic lupus erythematous (SLE), dermatitis herpetiformis, and rheumatoid arthritis. Pre-
liminary studies indicate that spleen extract may stimulate the immune system in conditions
such as HIV/ AIDS, leukemia, leprosy, Crohn's disease, and sickle cell disease. Some even
use spleen extract for glomerulonephritis, thrombocytopenia, ulcerative colitis, and a blood
vessel condition called vasculitis.
Some concern has been raised about the safety of spleen extract, as it is made of animal
spleens, which may be infected with some diseases: prion, bovine spongiform encephalitis, etc.
g) Thymus
The thymus (its name comes from the Greek word 9t)|i6c; and is called thymus because
its shape resembles that of a thyme leaf) is a specialized organ of the immune system. The
thymus "instructs" T- lymphocytes (T cells), which are critical cells of the adaptive immune
system. Each T cell attacks a foreign substance that it identifies with its receptor. 286 Thymus
extracts could have substances that influence the immune system, but it is very difficult to
exactly know what effect these extracts have on the immune system. The thymus seems to be
much less important in adults as it ceases to function after childhood.
The thymus is composed of two identical lobes and is located anatomically located in
the neck or chest of most vertebrate animals. In humans it is located in the anterior superior
285. Williams E. T. (1906), The Edibility of Animal Spleens, Reprinted from "American Medicine", Vol.
XI, No. 6, The Library of Congress, Washington DC.
286. Miller J. F. (2002). "The discovery of thymus function and of thymus-derived lymphocytes", Im-
munological Reviews, 185 (1): 7-14.
78
Animal by-products (ABPs): origins, uses, and European regulations
mediastinum, which is in front of the heart and behind the sternum. It is especially impor-
tant in newborn babies: without a thymus a baby's immune system collapses and the baby
will die. Histologically, the thymus can be divided into a central medulla and a peripheral
cortex that is surrounded by an outer capsule.
The functions of the thymus were not well known until a few years ago. During autop-
sies it was noticed that young adults that had died in traumatic accidents often had much
larger thymus glands than those dying from diseases of a chronic nature. The importance of
the thymus in the immune system was discovered by Miller 287 who surgically removed the
thymus from three day old mice and observed the subsequent deficiency in a lymphocyte
population. There are several immune substances in the body and it is difficult to predict all
the potential interactions when ingesting a thymus glandular. Furthermore, there are wide
variations in response between different people and different animals. 288
Thymus glands are available only from young animals (lambs and calves). The glands are
covered by a capsule of fibrous connective tissue that penetrates the gland and divide it into
lobules. The amount of connective tissue and fat increases with animal's age.
Thymus gland of neonatal calves is a source of many products. The thymus is responsible
for the production of T-lymphocytes, as well as the production of various hormones including
thymosin, thymopoeitin, thymulin, thymic humoral factor, and serum thymic factor.
Thymosin
Thymosin a 1 is believed to be a major component of Thymosin Fraction 5 289 respon-
sible for restoring immune function in animals lacking thymus glands. 290 Thymosin alpha 1
has been used for a number of years in cancer treatment 291 in order to increase the body's im-
mune system. It is now approved in 40 countries for the treatment of Hepatitis B and C. 292
Currently, a variety of other active ingredients of the thymus gland are being tested in
287. Miller, J. F. (2004), "Events that led to the discovery of T-cell development and function: A per-
sonal recollection", Tissue Antigens, 63 (6): 509-17.
288. For more notices see: Pearse G. (2006), "Normal Structure, Function and Histology of the Thy-
mus", Toxicologic Pathology, 34 (5): 504-514.
289. Thymosin fraction 5 contains several distinct hormonal-like factors which are effective in partially
or fully inducing and maintaining immune function (See: LowT. L., G. B. Thurman, C. Chincarini, J.
E. McClure, G. D. Marshall, S. K. Hu, A. L. Goldstein (1979), "Current status of thymosin research:
evidence for the existence of a family of thymic factors that control T-cell maturation", Annals of the
New York Academy of Sciences, 332: 33-48.
290. Garaci E. (2007), "Thymosin alpha 1: a historical overview", Annals of the New York Academy of
Sciences ("Thymosins in Health and Disease First International Symposium", Pages xi— xii, 1—468), vol.
1112: 14-20; Li J., C. Hui Liu, F. Shan Wang (2010), "Thymosin alpha 1: Biological activities, ap-
plications and genetic engineering production", Peptides, 31 (11): 2151-2158.
291. Garaci E., F. Pica, G. Rasi, C. Favalli (2000), "Thymosin alpha 1 in the treatment of cancer: from
basic research to clinical application", International Journal of Immunopharmacology, 22 (12): 1067—
1076; Schulof R. S., M. J. Lloyd, P. A. Cleary, S. R. Palaszynski, D. A. Mai, J. W. Cox Jr, O. Alabaster,
A. L. Goldstein (1985), "A randomized trial to evaluate the immunorestorative properties of synthetic
thymosin-alpha 1 in patients with lung cancer", Journal of 'Biological Response Modifiers, 4 (2): 147-158.
292. Garaci E., C. Favalli, F. Pica, P. Sinibaldi Vallebona, A. T. Palamara, C. Matteucci, P. Pierimarchi,
A. Serafino, A. Mastino, F. Bistoni, L. Romani, G. Rasi (2007), "Thymosin alpha 1: from bench to
bedside", Annals of the New York Academy of Sciences, 1112: 225-34.
79
Raffaella Leoci
terms of their efficacy, such as thymosine beta 4, thmostimuline, and others. Thymus pep-
tides are used for the treatment of immuno-deficiency diseases.
Thymus extract
Thymus extract 293 is used for infectious diseases including recurrent respiratory infec-
tions, colds, flu, H1N1 "swine" flu, hepatitis B, hepatitis C, Epstein-Barr virus (EBV),
mononucleosis, herpes and shingles, sinusitis, and AIDS/HIV. It is also used for asthma,
hay fever, food allergies, cancer, rheumatoid arthritis (RA), chronic fatigue syndrome (CFS),
and systemic lupus erythematosus (SLE). Other uses include maintaining white cell produc-
tion in cancer patients treated with radiation or chemotherapy and preventing the effects
of aging. An increasing number of manufacturers offer thymus extracts for various diseases.
Thymus is also used in cosmetics to possibly slow skin aging processes and in hair care
products because of its film-forming properties and peptides and amino acids.
For foods
The thymus glands from lamb and calf are blanched to firm the tissue and peeled from
the capsule before they are cooked by frying or stewing.
When animal thymic tissue is sold in a butcher shop or at a meat counter, thymus is
known as sweetbread. 294
h) Thyroid
All vertebrates have a thyroid gland that is extremely vascular. 295 In mammals, it is usu-
ally bilobed and located just caudal to the larynx and adjacent to the lateral surface of the
trachea. The two lobes may be connected by a fibrous isthmus (e. g., ruminants, horses, etc.),
or a connecting isthmus may be indistinct (e.g. dogs, cats). 296 This gland produces, stores,
and secretes the thyroid hormones that are the only iodinated organic compound in the
body necessary for growth and proper metabolism.
Thyroid tissue is composed of millions of tiny saclike follicles, which store thyroid hor-
mone, namely thyroglobulin, a glycoprotein. Blood capillaries attached to the gland yield a
constant supply of plasma. Thyroglobulin is the chief component of the colloid, a jellylike
substance that is secreted by the follicles. It attaches to the thyroid hormone for storage
purposes. Before it is released into the bloodstream, the thyroid hormone is converted into
293. Thymus extract is a substance that is collected from the thymus gland of a cow, but it also can
be manmade. The extract is believed to produce multiple health benefits, including boosting the hu-
man immune system. The use of thymus extract is known to posses certain risks. Since the extract is
collected from the thymus gland of cows, there is a risk of contamination when the extract is collected
from a diseased animal: the infection may transfer to humans.
294. "Sweetbread" is the name of a dish made of the animal's brain, pancreas, or thymus gland (neck/
throat/gullet sweetbread) of an animal younger than one year old.
295. Among others see: Stathatos N. (2006), Anatomy and Physiology of the Thyroid Gland, in Thyroid
Cancer. A Comprehensive Guide to Clinical Management, L. Wartofsky, D. Van Nostrand (Edts.), 2nd
ed., 2006, Humana Press, Totowa, New Jersey (USA), pp. 3-7.
296. Reece W. O. (2009), Functional Anatomy and Physiology of Domestic Animals, 4° Edition, Wiley-
Blackwell, Ames, Iowa (USA), pp. 165-7; Swindle M. M. (1998), "Comparative anatomy and physiol-
ogy of the pig", Scandinavian Journal of Laboratory Animal Science, 25 (1): 1 1-21.
80
Animal by-products (ABPs): origins, uses, and European regulations
thyroxine 297 and small quantities of the other thyroid hormones. The quantity of thyroxine
production depends on a sufficient intake of iodine and on stimulation by the thyroid-
stimulating hormone (TSH) sent from the pituitary gland. Secretion of thyroid hormones,
mostly thyroxine (T4), is controlled by thyroid-stimulating hormone (TSH), which is re-
leased by the pituitary gland when the level of thyroid hormones in the blood drops below
a certain threshold.
In other words, the thyroid gland produces the hormone thyroxine, known as T4. The
gland also secretes 3,5,3'-triiodothyronine (known as T3), and other deiodinated metabolites.
T3 is about 3-5 times more potent than T4, while reverse T3 is thyromimetically inactive.
In conclusion, the body, when functioning properly, converts some of its thyroxine (T4)
to triiodothyronine (T3), which is the major active thyroid hormone. Other hormones called
T2 and Tl are also formed, although their precise functions are not yet fully understood. In the
bodies, about 80% of triiodothyronine derive from conversion of thyroxine, a process medi-
ated by deiodinase enzymes, the remainder coming directly from the thyroid gland.
Metabolic disorders happen when the thyroid secretes too little (hypothyroidism) 298 or
too much thyroxine (hyperthyroidism). The first occurs when there is insufficient iodine in
the diet. Excessive secretion of thyroxine known as myxedema in the adult and cretinism in
infancy and childhood, comes from glandular malfunction.
The thyroid gland also produces the hormone calcitonin, a compound that stimulates
deposition of calcium from the blood into the bones, balancing the action of parathyroid
hormone.
Thyroid extracts
The treatment of hypothyroidism is to administer of thyroxine. Natural thyroid extracts
have been used since 1891, when Murray 299 used an extract, made from dried animal glands
to treating of myxedema. This use was approved in USA by the FDA in 1939.
Desiccated thyroid extract was the most common form of treatment for hypothyroidism
until the discovery of the individual thyroid hormones (T3 and T4) and their commercial
297. Tyroxine is a precursor of thyroid hormone and the neurotransmitters dopamine, norepinephrine,
and epinephrine. A deficiency of tyrosine leads to hypothyroidism and low adrenal function. The rec-
ommended daily amount of tyroxine is about 1 g/day for adults.
298. Some disorders related to thyroid deficiency are depression, poor concentration, memory distur-
bances, cold hands and feet, accumulation of excess body fat, dry skin, difficulty in losing weight, men-
strual problems, thin hair, elevated cholesterol, migraine headaches, infertility and hypertension (See:
Stanosz S. (1992), "Levels of thyroid hormones and thyrotropic hormone in serum of women with peri-
menopausal arterial hypertension", Ginekologia Polska, 63 (3): 130-3; Saito I., K. Ito, T. Saruta (1983),
"Hypothyroidism as a cause of hypertension", Hypertension, 5: 112-115; MoreauT. (1988), "Headache
in hypothyroidism. Prevalence and outcome under thyroid hormone therapy", Cephalalgia, 18: 687-9;
Silva J. E. (2001), "The multiple contributions of thyroid hormone to heat production", The Journal of
Clinical Investigation, 108 (1): 35-37; Fazio S., E. A. Palmieri, G. Lombardi, B. Biondi (2004), "Effects
of Thyroid Hormone on the Cardiovascular System", Endocrine Reviews, 59 (1): 31-50; Vierhapper H.
H. (1997), "Assessment of thyroid gland function in unwanted infertility—indications forTRH test and
clinical impact from the viewpoint of the endocrinologist", Acta Medica Austriaca, 24 (4): 133-5; Hey-
mann W. R. (2008), Thyroid Disorders with Cutaneous Manifestations, Springer- Verlag, London).
299. Murray G. R. (1891), "Note on the treatment of myxoedema by hypodermic injections of an
extract of the thyroid gland of a sheep", British Medical Journal, 2: 796-7.
81
Raffaella Leoci
availability. During the 1960s, science-based physicians stopped using desiccated thyroid ex-
tract because its potency varied from batch to batch, which caused difficulty optimizing the
patient's thyroid hormone levels. In the early 1960s, the development of synthetic thyroid
hormones overshadowed animal hormones in doctor approval. Since then, animal thyroid
hormones have taken a back seat to synthetic hormones. Today, desiccated thyroid 300 is made
from pig thyroid glands.
Currently the opinions on the use of the two products (natural thyroid extracts and
synthetic thyroid hormones i.e. thyroxine-T4) are different because they have both strengths
and weaknesses.
The UK Royal College of Physicians has recommended the exclusive use of synthetic
levothyroxine for the treatment of hypothyroidism. Synthetic pills contain a pure form of
the hormones produced in the body, as it consist solely of T4. While they are called "syn-
thetic", these products are actually identical to the natural compound T4.
At the present time, at least in Europe, a large majority of people of all ages who have
hypothyroidism are being treated with levothyroxine.
.In animal-originated thyroid pills, the amounts of T4 andT3 vary among manufac-
turers. Moreover, the preparations from any single manufacturer over time will also vary. A
perfect dose one month may be too little or too much a couple of months later, when the
next batch comes on the market. It follows that desiccated thyroid extracts are not as precise
as synthetic compounds. In addition, it is difficult to prescribe the ideal hormone dosage.
Furthermore the hormone balances in animals are not the same as in humans, so it is not
in the human's best interest to argue that animal thyroid hormones are truly natural. Many
doubts surround the idea that tissue from an animal thyroid gland will migrate to the same
organ site in the human body when ingested, and the idea assumes that digestion would not
destroy the tissue. Animal thyroid pills aren't purified, which means that they contain sub-
stances that aren't naturally found in humans. Researchers have yet to determine how these
substances affect the human body.
Finally, it should be remembered that in 2005, FDA enforcement removed more than
half a million bottles of "Armour Thyroid" from US pharmacies due to unstable concentra-
tions of thyroid hormone in the preparation.
Opposing opinion regarding the use of thyroid extract
The advocates of the use of animal thyroid extract have an opposing opinion. According
to them thyroid extracts made from a healthy thyroid gland are better than synthetic thy-
roxine because they contain both T4 and T3 and unmeasured amounts of diiodothyronine
(T2), monoiodothyronine (Tl), calcitonin, and other protein-bound iodine.
Natural thyroid extracts containing T4, T3, and Tl, T2, even if they derive from ani-
mals (pig, cattle, etc.), are identical in molecular structure to the human hormones. There-
fore, there are many advantages in using them. Instead, synthetic preparations consist solely
of T4 and depend on the body to convert the T4 to T3 and other metabolites. For those peo-
300. Desiccated thyroid is the dried and powdered thyroid gland. During the process of preparing
this glandular, the fat and connective tissue are removed. Desiccated thyroid is often from hogs, but
may also from cows and sheep. The pharmaceutical preparation is standardized and contains both
thyroxine and triiodothyronine. "Armour Thyroid" or "Nature-Throid" & "Westhroid" the name of a
commercial brand in USA.
82
Animal by-products (ABPs): origins, uses, and European regulations
pie whose problem is not a decline in T4 production but a defect in the conversion of T4 to
T3, taking T4 may be useless. 301 Moreover, the various thyroid hormones surely exist in the
body for a reason, so supplementing with the thyroid hormone spectrum seems necessary.
Harrower 302 was one of the leading advocates of the "Organotherapy" approach. 303 Ac-
cording to him, "clinical experience has established this beyond all doubt - literally thou-
sands of cases of dyscrinism 304 having been treated with single endocrine products without
satisfactory results have, on changing to an indicated pluriglandular remedy, shown results
so different that they are remarkable."
It should be noted that even the effect of soy on thyroid function is currently a contro-
versial topic. Some believe that soy increases metabolic rate and thyroid function, others say
it has no effect. 305 Several recent articles, however, have noted problems with people taking
both soy and thyroid supplements at the same time. 306
Divi and others 307 have identified the mechanism of soy's effect on thyroid function.
Genistein and daidzein, the isoflavones in soy, inhibited thyroid peroxidase by acting as
alternative substrates. They conclude that "because inhibition of thyroid hormone synthesis
can induce goiter and thyroid neoplasia in rodents, delineation of anti-thyroid mechanisms
for soy isoflavones may be important for extrapolating goitrogenic hazards identified in
chronic rodent bioassays to humans consuming soy products."
4.7 Hides and skins
The largest components, based on value and volume, of ABPs derived from the slaughter
of food animals are the hides and the skins, 308 in particular those derived from cattle. Animal
hides have been used for shelters, clothing and as containers (water, foods, etc.) since prehis-
toric times. Hides and skins are generally one of the most valuable by-products from animals.
301. Bunevicius R., G. Kazanavicius, R. Zalinkeviiius, A. J. Prange Jr. (1999), "Effects of thyroxine
as compared with thyroxine plus triiodothyronine in patients with hypothyroidism", New England
Journal of Medicine, 340 (6): 424-9.
302. Harrower H. R. (1922), Practical organotherapy: the internal secretions in general practice, The Har-
rower Laboratory, Glendale CA, p. 34.
303. Harrower H. R. (1914), Practical Hormone Therapy: A Manual of Organotherapy for General
Practitioners, Hueber Verlag GmbH & Co K, Ismaning (D).
304. It is a term used for any endocrine malfunction.
305. Dillingham B. L., B. L. McVeigh, J.W. Lampe, A.M. Duncan (2007), "Soy protein isolates of
varied isoflavone content do not influence serum thyroid hormones in healthy young men", Thyroid,
17 (2): 131-137.
306. Messina M., G. Redmond (2006), "Effects of Soy Protein and Soybean Isoflavones on Thyroid
Function in Healthy Adults and Hypothyroid Patients: A Review of the Relevant Literature", Thyroid,
16 (3): 249-258; Setchell K. D. (1998), "Phytoestrogens: the biochemistry, physiology, and implica-
tions for human health of soy isoflavones", American Journal of Clinical Nutrition, 68: 1333S-46S;
Conrad S. C, H. Chiu, B. L. Silverman (2004), "Soy formula complicates management of congenital
hypothyroidism", Archives of Disease in Childhood, 89 (1): 37-40.
307. Divi R. L., H. C. Chang, D. R. Doerge (1997), "Anti-Thyroid Isoflavones from Soybean: Isola-
tion, Characterization, and Mechanisms of Action", Biochemical Pharmacology, 54 (10): 1087-1096.
308. Wordreference is implying that both Hide and Skin are synonyms, having roughly the same
meaning. But in the current language "hides" and "skins" are two different parts of the same animal:
the difference is that hides are thicker: we would say cowhide and pigskin.
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Raffaella Leoci
The hides represent a great portion of the weight of the live animal: from 4% to as much
as 1 1%. 309 After the hide or skin is removed from the animal, it should be cured 310 quickly
to avoid decomposition by bacteria and enzymes.
There are four basic treatments: airdrying, curing with salt, curing by mixer and curing
by raceway. 311 Some skins, such as sheepskins and pigskins, contain a large quantity of fat. It
is desirable to reduce the fat to approximately 3.0% on a dry-weight basis. It takes four weeks
to process raw hides into leather.
The skin of virtually every animal can be used to produce leather. Animal skins have
been the source of clothing attire for man since prehistoric times. Leather is used in a re-
markable number of applications, including automobile and furniture upholstery, shoes,
sporting goods, luggage, garments, gloves, and purses. A representative of the leather indus-
try categorized leather utilization as 40% for upholstery, 50% for shoes and shoe leather, and
10% for other uses.
Slaughterhouse skin by-products can be treated and become a good protein source. The
skin is shredded into little pieces and washed. The wash water is filtrated for impurities and
the dissolved proteins are concentrated. These concentrated proteins are mixed back into
the clean and centrifuged skin-shreds; the skin-shreds absorb the concentrated protein and
can be dried. The dry material will contain approx. 7% of water and have an acceptable
industrial shelf life.
The skin of some animal species is also used for processed meat products. This is the
case with pork skin and poultry skin, in some cases also with calf skin, particularly from calf
heads and legs. 312
Gelatin
Animal gelatin 313 is an edible jelly produced by the controlled hydrolysis of a water-insolu-
ble collagen derived from proteins extracted from fresh animal tissues (mainly skins and bones)
that have arrived in an edible condition, through boiling. Both hides and bones contain large
quantities of collagen. Commercially available gelatin is a dry powder of various granule sizes.
Mixed with water the protein molecules of the gelatin absorb water and form a gel.
The processing of gelatin from hide consists of three major steps: elimination of non
collagenous material from the raw material, controlled hydrolysis of collagen to gelatin,
recovery and drying of the final product.
309. E.g. cattle: 5.1 - 8.5%, average: 7.0%; sheep: 11.0 - 11.7%; swine: 3.0 - 8.0%.
310. Long ago, the Native Americans used wood, woodsmoke and animal brain tissue as tanning
agents to cure and preserve animal hides.
311. Anderson R. M. (1948), Methods of Collecting and Preserving Vertebrate Animals, National Mu-
seum of Canada, Bulletin No. 69, Ottawa; O'Flaherty E, W. T. Roddy, R. Miller Lollar (1978), The
chemistry and technology of leather, R. E. Krieger Pub. Co., Huntington, N.Y; Thorstensen T. C. (1985),
Practical leather technology, R. E. Krieger Pub. Co., Huntington, N.Y; Minnoch J. K., S. R. Minnoch
(1979), Hides and skins, National Hide Association, Chicago 6, Ill-USA.
312. Chicharron (Spanish, Portuguese: Torresmo, Filipino: Chicharon) is a food made of fried pork
rinds. It is sometimes made from chicken, mutton, or beef.
313. Lambert T. (2008), Glue, Gelatine and Their Allied Products: A Practical Handbook for the
Manufacturer & Agriculturist, BiblioBazaar, Charleston (USA); The Manufacture of Glue and Gela-
tine; the Application and Uses of Machinery, Etc. New York, Chicago, etc., National Provisioner
Publishing Co. General Books, 2010.
84
Animal by-products (ABPs): origins, uses, and European regulations
Animal gelatin is either spray dried as simple gelatin or as a hydrolyzed gelatin, i.e. partial
or total splitting of the long gelatin molecules into smaller fragments. This is done with chemi-
cal or enzymatic processes followed by refining, and other unit operations, to obtain the re-
quired product cleanness and or property. To ensure an economically feasible drying process, all
gelatin products are concentrated before drying. After being concentrated, the gelatin is dried
by one of many methods, such as being cooled in a drying tunnel, drum drying or spray drying.
Another extraction procedure for gelatin is acid processing. This is usually applied to
pig skin or bone.
Gelatins are used in the food and pharmaceutical industries. When it is used in food in-
dustries is added to a wide range of foods, forming a major ingredient in jellies and aspic. Its
main use is the production of jellied desserts, because of its "melt in the mouth" properties,
and it is is also added to a range of meat products, i. e. to meat pies. Gelatins are also widely
used as a stabilizer for ice cream and other frozen desserts. They are added as a protective
colloid to ice cream, yoghurt and cream pies. The raw material can also be rendered into lard.
Collagen from hides and skins has a role as an emulsifier in meat products because it can
bind large quantities of fat. This makes gelatins a useful additive or filler for meat products.
In USA and in Europe the pharmaceutical industry uses about 6.5% of the total produc-
tion of gelatin. Most of it is used to make the outer covering of capsules and as a binding and
compounding agent in the manufacture of medicated tablets and pastilles. It is used as ingredi-
ent in protective ointment, such as zinc gelatin for the treatment of ulcerated varicose veins.
Since gelatin is a protein, it is used as a plasma expander for blood in cases of very severe
shock and injury. A product made from extracted collagen can stimulate blood clotting dur-
ing surgery. Pork skin is similar to human skin, and can be converted into a dressing for burns
or skin-ulcers. Prior to use, pork skin used as a dressing needs to be cut into strips or into a
patch, shaved of hair, split to a thickness of 0.2 -0.5 mm, cleansed, sanitized and packaged.
Gelatin is an excellent emulsifier and stabilizing agent for many emulsions and foams. It
is used in cosmetic products, in silk screen printing inks, and photogravure printing, etc. 314
Pork skin
It is normally used as food, unlike other animal skins that are used in leather produc-
tion. Nonetheless, pork skin is a sometimes used as a fabric for footwear and garments. Be-
cause it is similar to human skin, it can be converted into a dressing for burns or skin-ulcers.
Pork skin is collagen rich and in precooked form a valuable material for the manufac-
ture of some meat products of the precooked-cooked variety. Occasionally raw pork skin is
shredded into small particles and used in processed meat products . It can also be used for
gelatin production.
Collagen, that is a fibrous protein abundant in all animals, is used in the food industry
as casing and wraps. A wrapping film for hams and other food products can be made using
collagen membranes from pig skin.
Skin glue manufacturing uses the parings and cuttings of hides from tan-yards: for
example the ears of oxen and sheep, and the skins of rabbits, hares, cats, dogs and other
animals. Supposedly the best skin glue is obtained from a mixture of the hide, ear and face
clippings of the ox and calf. For art applications, rabbit, sheep and deer skin glue are popular.
314. GMIA, Gelatin Manufacturers Institute of America, Gelatin Handbook, Written and produced by
the members of the GMIA, 2012.
85
Raffaella Leoci
Rabbit skin
Rabbit skin glue is made from rabbit skin, tendon, bone and gut and is used in making
and repairing instruments, traditional woodworking, gilding and painting techniques.
Chicken skin
Chicken skin is removed from the carcass or from individual cuts. It has a high fat
content and is ground prior to being added to processed meat products. Chicken fat serves
as the fat portion in all-chicken processed meat products (chicken frankfurters, chicken
nuggets, and chicken bologna). Chicken skin is added to meat products for the same
purpose as pork fat in pork/beef products, namely to contribute to product flavor and
produce a soft texture.
4.8 Hairs and wools, nails, horns, feathers, hooves
Keratin
Keratin is one of the main components of these products. The earliest documented
use of keratin in medicine comes from a Chinese herbalist named Li Shi-Zhen in the Ming
Dynasty, as recalled by Ben Cao Gang Mu. 315 Around 1849, the word "keratin" appears in
the literature to describe the material that made up hard tissues such as animal horns and
hooves. In the following years many scientists were more interested in dissolving hair and
horns in order to make better products. In the years preceding World War I, lime was applied
to the manufacture of keratin gels. 316 From 1900 to 1940, several methods were developed
to obtain keratins using oxidative and reductive chemistries. 317 Among the first inventions
were keratin powders for cosmetics, composites, and coatings for drugs, respectively. 318 It
was during the years of World War II and after that one of the most comprehensive research
projects on the structure and chemistry of hair fibers was undertaken: between the years of
1940 and 1970, nowhere in the world was keratin research more active than in Japan. Dur-
ing the same years in Australia, the Council for Scientific and Industrial Research (later the
Commonwealth Scientific and Industrial Research Organisation or CSIRO) was founded
in order to better understand the structure and chemistry of wool fibers. Researchers at
CSIRO conducted many of the most fundamental studies on the structure and composition
315. Ben Cao Gang Mu 0 ), Materia Medica, a dictionary of Chinese herbs, written by Li Shi
Zhen (^Sf#) (1518-1593). It consists of 52 volumes. The booklists 1,892 medical material of herbs,
animals and mineral with 1 1,096 formulae being used in the past. The book has been translated into
more than 60 languages (Japanese, French, German, Russian, Latin, etc.).
316. In a German patent issued in 1905 (n. 184,915), John Hoffmeier described a process for extract-
ing keratins from animal horns using lime.
317. Brein F, O. Baudisch (1907), "The oxidative breaking up of keratin through treatment with
hydrogen peroxide", Zeitschrift fur Physiologische Chemie, 52: 158-69; Neuberg C, Process of producing
digestible substances from keratin, US pat. n. 926,999, 1909; Goddard D. R., L. Michaelis (1934), "A
study on keratin", The Journal of Biological Chemistry, 106: 605-14.
318. Beyer C, The keratin or horny substance of the hair, German pat. N. 22,643, October 14, 1907;
Goldsmith B. B., Thermoplastic composition containing keratin, US pat. N. 922692, May 25,1909; Dale
H. N. (1932), "Keratin and other coatings for pills", Pharmaceutical Journal, 129: 494-5.
86
Animal by-products (ABPs): origins, uses, and European regulations
of wool. In 1965, CSIRO scientist Crewther and others 319 published the definitive text on
the chemistry of keratins.
During the 1980s, a number of scientists began to explore the potential uses of keratins.
In 1982, Japanese scientists 320 published the first study describing the use of a keratin coating
on vascular grafts as a way to eliminate blood clotting, as well as experiments on the bio-
compatibility of keratins. 321 Soon thereafter in 1985, two researchers from the UK published
a review article speculating on the prospect of using keratin as the building block for new
biomaterials development. 322 A review by Rouse and Van Dyke 323 stated that extracted kerat-
ins were capable of forming self- assembled structures that regulated cellular recognition and
behavior. The studies and discovered qualities led to the development of keratin biomaterials
with applications in wound healing, drug delivery, tissue engineering, trauma treatment and
medical devices manufacturing.
In addition, these and others studies concluded that keratins were a group of strong
fibrous, structural proteins and were the key structural material making up the outer layer
of human and animal skin. Keratins are also the key structural component of hair and nails.
Keratin amino acids have several unique properties and, depending on the levels of the vari-
ous amino acids, they can be inflexible and hard like hooves, or soft as is the case with skin.
The polypeptide chains of keratin are arranged in parallel sheets held together by hydrogen
bonding. Approximately 10 keratins form the basis of hair or claw, with a further 20 found
in internal body cavity epithelia. 324 In humans 54 functional keratin genes exist. 325 Cysteine
is a major component of keratin proteins; it is an attractive material for the absorption of
heavy metals from aqueous solution.
Keratin compounds can be hydrolyzed to form many useful materials and numerous
patents, especially in USA and Japan. 326 have been issued.
319. Crewther W. G., R. D. B. Fraser, F. G. Lennox, H. Lindley (1965), The Chemistry of Keratins, in
C. B. Anfinsen Jr., M. L. Anson, J. T. Edsall, and F. M. Richards (Edts) , Advances in protein chemistry,
Academic Press, New York, pp. 191-346.
320. Noishiki Y., H. Ito, T. Miyamoto, H. Inagaki (1982), "Application of denatured wool keratin
derivatives to an antithrombogenic biomaterial: Vascular graft coated with a heparinised keratin de-
rivative", Kobunshi Ronhunshu, 39 (4): 221-7.
321. Ito H., T. Miyamoto, H. Inagaki, Y. Noishiki (1982), "Biocompatibility of denatured keratins
from wool", Kobunshi Ronhunshu, 39 (4): 249-56.
322. Jarman T., J. Light (1985), "Prospects for novel biomaterials development", World Biotechnology
Rep, vol. 1, pp. 505-12.
323. Rouse J. G., M. E. Van Dyke (2010), "A Review of Keratin- Based Biomaterials for Biomedical
Applications", Materials, 3: 999-1014.
324. The first to point out that there are several types of keratins was Lissizin T. (1928) ("The oxida-
tion products of keratin by oxidation with permanganate II", Zeitschrift fur Physiologische Chemie, 173:
309-11).
325. Moll R., M. Divo, L. Langbein (2008), "The human keratins: biology and pathology", Histochem-
istry and Cell Biology, 129 (6): 705-733.
326. Among the most interesting: US Patents n. 3,642,498 (1972); 4,240,450 (1980); 4570629
(1986); 5047249 (1991); 6,270,791 (2001); 6,316,598 (2001); 6,270,793 (2001); 6,274,155 (2001);
6.461,628 (2001). Japan Patents Appl. n. S62-333838 (1989); 3-223207 (1991) 04082561 (1992);
247925 (2008); 023924 (2009).
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Raffaella Leoci
Feathers
Internationally, feathers production is huge. 327 Feathers are used in a number of decora-
tive products, such as boas, feather fans, masks, costume accessories, bird ornaments and
even earrings and flowers. Down and feathers are used for insulation and padding of prod-
ucts like jackets, bedding, sleeping bags, and pillows. Most of these products contain a blend
of down and feathers. The production and harvesting of fine feathers and down for use in the
garment and household linen industries is different than the processing of coarse feathers for
feather meal. In the USA much of the production of coarse product is converted into feather
meal that is hydrolyzed for use in animal feed supplements, fertilizers and other products.
Since post World War II, hundreds of patents have been issued for these uses (insulating
products, fiber products, oil coagulants, pest repellent, etc.). Dalev 328 developed and pro-
moted a combined enzyme-alkaline technology for processing waste feathers from poultry
slaughterhouses. On the other hand, Kumar and others 329 have experimented with the use of
a type of bacillus to biodegrade poultry feathers.
Horns and hooves
Horns and hooves are used as raw material for fertilizers that are excellent source of slow
release nitrogen; for the production of protein based fire-fighting agents; for making foam-
ing agents; and for light weight cellular concrete additives.
Wool and hair
Wool and hair have multiple uses based on their fiber properties. These qualities guide
their usage into fabric, building insulation, and absorptive products. Synthetically derived
products have challenged hide, skins, wool, and hair in nearly all traditional applications and
will undoubtedly continue to do so in the future.
Fire fighting foams
Hooves, horns, and feathers can be useful to make fire fighting foams. Fire fighting
foams are a collection of bubbles formed by the aeration of a foam concentrate solution with
water. It follows that foams are made up of three components: foam concentrate (a liquid
327. World-wide poultry processing plants are producing millions of tons of feathers as waste products
annually. Feathers represent 5-7% of the total weight of mature chickens. Around 24 billion chickens
are being killed per year across the world which is discarding about 2 million tonnes of poultry feather.
Feather is generated in bulk quantities as a by-product of poultry industry. Typically as each bird has
up to 125gms of feather, the year worldwide production of feather waste is about 155.000 tons. As
regards the geese, they moult the natal plumage into juvenile feathers between 3-5 weeks of age and
than moult that juvenile plumage into adult plumage between 8-11 weeks of age. Feather weight of
an adult goose makes up about 6.2% of its total body weight (See: Kozak J. (201 1), "An Overview of
Feathers Formation, Moults and Down Production in Geese", Asian-Australasian Journal of Animal
Sciences, 24 (6): 881-887).
328. Dalev P. G. (1994), "Utilisation of waste feathers from poultry slaughter for production of a
protein concentrate", Bioresource Technology, 48 (3): 265-267.
329. Kumar E. Vijay, M. Srijana, K. Chaitanya, Y. H. Kumar Reddy, G. Reddy (2011), "Biodegra-
dation of poultry feathers by a novel bacterial isolate Bacillus altitudinis GVC1 1", Indian Journal of
Biotechnology, 10: 502-507.
88
Animal by-products (ABPs): origins, uses, and European regulations
produced by chemical manufacturers and supplied in drums or bulk), water and air. Gener-
ally, the task of fire fighting is to cool the fire and to coat the fuel, preventing its contact with
oxygen, in order to suppress the combustion. 330 Fire-fighting foam, floats on a fuel surface
separating it from oxygen. Other components of fire-retardant foams are organic solvents,
foam stabilizers, and corrosion inhibitors.
Over the years, foams have been classified in different ways. The earliest foams were
based upon a chemical reaction occurring between aluminum sulphate and sodium bicar-
bonate. This type of foam is now largely obsolete. Nowadays there are two types of foams:
synthetic and protein foams.
The first type is made, commonly, from a mixture of diammonium sulphate, diammonium
phosphate, monoammonium phosphate, besides gum thickeners, an iron oxide coloring agent,
and preservatives: 331 for long-term fire retardants fertilizer salts, are typically mixed in with
water to ensure uniform dispersal. Even after the water has evaporated from these synthetic
foams, the retardant remains effective until it is removed by rain or erosion. 332 The foams form
a combustion barrier after the evaporation of the water carrier, and its effectiveness depends on
the amount of retardant per unit surface area. There are other types of synthetic foam forms
based on mixtures of non-fluorochemical, hydrocarbon type surfactants along with solvents
and water. These foam agents do not form aqueous films or polymeric membranes. Instead,
they function by forming an aggregate of foam bubbles on the surface of the fuel.
Protein foams were the first types of mechanical foam to be marketed extensively and
have been used since World War II. 333 These foams are produced by the hydrolysis of gran-
ulized keratin 334 protein (protein hydrolysate) such as hoof and horn meal, chicken feathers,
etc. They produce a homogeneous, stable foam blanket that has excellent heat resistance,
burnback, and drainage characteristics. All contain surfactants, foaming, and wetting agents.
Stabilizing additives and inhibitors are included to prevent corrosion, resist bacterial decom-
position and to control viscosity. Fire retardants depend on the water they contain to retard
or suppress the fire. The foaming agents affect the rate at which water drains from the foam
and how well it adheres to the fuel. The surfactants and wetting agents increase the ability of
the drained water to penetrate fuels thus reducing their ability to ignite. Fuels are insulated
from heat and air contact is reduced. These retardants lose their effectiveness once the water
has evaporated or drained from them.
330. NF-National Foam, A Firefighter's Guide to Foam (www.Kidde-Fire.com); "Ideas Advance Fire
Fighting", Industrial Fire World Magazine, 2005, 20 (6): 10-12, 36-37.
331. Hamilton S., D. Larson, S. Finger, B. Poulton, N. Vyas, E. Hill (1998), Ecological effects of fire
retardant chemicals and fire suppressant foams, Northern Prairie Wildlife Research Center Home Page,
Jamestown, (ND-USA).
332. U.S. Department of Agriculture, Fire fighting chemical products - Differences and similarities,
Wildland Fire Chemical Systems, Missoula Technology and Development Center. U.S. Forest Service.
Missoula, (MT-USA) 2002. (http://www.fs.fed.us/rm/fire/Wildland_chemicals.html).
333. On the use of hooves and horns, to produce foams, have been issued numerous patents. One of
the newest is the Patent Family Members CN 101491723 A, Protein foam fire extinguisher produc-
tion method using hoof and horn grain (29-Jul-2009). The product of this invention seems to have
the advantages of long storage period, high foam expansion rate, less sediment, long drainage time and
high extinguishing efficiency.
334. The keratin helps to bond the foam bubbles into a durable blanket, which stops it breaking up on
impact with the fire and makes it very effective at smothering flames.
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Raffaella Leoci
Unlike synthetic foams, protein foams are bio-degradable, flow and spread slower, and
provide a foam blanket that is more heat-resistant and more durable. They have slow knock-
down characteristics but provide superior post fire security at very economical cost. They
may be used with fresh or seawater. The foam is specifically designed to quell the hotter,
high intensity fires triggered by aviation fuel and is therefore used to make a special fire ex-
tinguishing foam used by airport fire and rescue teams. Protein foams are especially intended
for use on hydrocarbon fuels.
There is significant potential for damage to terrestrial vegetation and to aquatic ecosys-
tems from synthetic fire fighting foams. The environmental effects of fire retardants have
been of concern since the 1970s. 335 However, few studies into the ecological effects of fire
retardants or foams on vegetation have been carried out. 336 Because the protein foams are
biodegradable, the damage on the environment should be more limited.
4.9 Hearts
Heart is used as a table meat: it can be roasted, braised, grilled. Heart meat is often also
used as an ingredient in processed meats. It can be also used for transplantation.
Xenotransplantation (XT) is the transplant of organs, cells, or tissues between species.
It commonly refers to transplants of animal organs (e.g., nonhuman primates or pigs) into
humans. 337
Xenotransplantation is not a recent phenomenon. 338 Researchers have made sporadic
attempts at cross-species transplants from early as the 17th century with little success. Pri-
mate donors were used in early whole organ experiments. Pig heart valves have been used
in humans for several decades; however they are first chemically treated to kill pig cells. 335
In 1999, the U.S. FDA 340 banned clinical trials with non-human primates. On April 6th,
335. Dodge M. (1970), "Nitrate poisoning, fire retardants, and fertilizers-any connection?", Journal of
Range Management, 23 (2): 44-247.
336. Between these: Gaikowski M. P., S. J. Hamilton, K. J. Buhl, S. F. McDonald, C. H. Sum-
mers (1996), "Acute toxicity of three fire-retardant and two fire-suppressant foam formulations to the
early life stages of rainbow trout (Oncorhynchus mykiss)", Environmental Toxicology and Chemistry,
15 (8): 1365-1374; McDonald S. E, S. J. Hamilton, K. J. Buhl (1996), "Acute toxicity of fire control
chemicals to Daphnia magna (Straus) and Selanastrum capricornutum (Printz)", Ecotoxicology and
Environmental Safety, 33: 62-72; McDonald S. E, S. J. Hamilton, K. J. Buhl (1997), "Acute toxicity of
fire-retardant and foamsuppressant chemicals to Hyalella azteca (Saussure)", Environmental Toxicology
and Chemistry, 16 (7): 1370-1376; Ecological Risk Assessment: Wildland Fire-Fighting Chemicals,
Prepared by LABAT Environmental for Missoula Technology and Development Center, USDA Forest
Service, Missoula, (MT-UA) 2007.
337. Buhler L. H. (Edtr.) (2013), Xenotransplantation, John Wiley & Sons A/S, London.
338. Kttss R., P. Bourget (1992), Une histoire illustree de la greffe d'organes. La grande aventure du siecle,
Laboratoires Sandoz, Rueil-Malmaison (France); Deschamps J. -Yves, F. A. Roux, P. Sal', E. Gouin
(2005), "History of xenotransplantation", Xenotransplantation, 12 (2): 91-109; Barber N. (2007), Or-
gan Transplanting. The Cannibalistic Nature of Transplant Medicine, Third Edition, Adelaide, Australia,
(standardoil@hotmail.com).
339. For a detailed chart of all xeno experiments using cells or organs from non-human primates see
the Council of Europe's July 2000 report on xenotransplantation (Tallacchini M., Existing regulations
on xenotransplants in the European area (COE, EU and EEA countries), XENOME WPS, D274).
340. Butler D. (1999), "FDA warns on primate xenotransplants", Nature, 398 (6728): 549-637.
90
Animal by-products (ABPs): origins, uses, and European regulations
1999, the U.S. FDA stopped short of banning nonhuman primate xenografts, encourag-
ing "further scientific research" into the risks. The European legal aspects are discussed by
Taupitz and Weschka. 341
Since the mid-1990s the European Institutions and the European Agency for the Evalu-
ation of Medicinal Products (EMEA) adopted diverse xenotransplantation policies. In 1997,
the Council of Europe (COE) approved Recommendation 15/97 on the necessity to set
standards for xenotransplantation clinical trials. At that time, the European countries - those
belonging to the Council of Europe, to the European Union (still European Communities at
that time), and to the European Economic Area (EEA) - began taking steps to regulate XT.
In 1999, the Parliamentary Assembly of COE asked the Member States to take into
account a moratorium on all clinical trials with XT, and some countries (such as Denmark,
The Netherlands, and Norway) agreed; others set up dedicated institutions (UK) or technol-
ogy assessment procedures (Switzerland) to improve control or understanding of the impli-
cations of XT.
In 2007, the approval of Regulation 2007/1394/EC has provided a direct regulation for
xeno-cell and tissue-based products, having established a central approval procedure for the
market placing of so-called advanced therapies.
From 1997 until 2009, the EU xenotransplantation policy was not subject to as fierce
a public dispute and scrutiny at the European level as other areas of biotechnology and bio-
medicine. The European Council briefly discussed xenotransplantation in the context of the
clinical trials Directive. On 25 September 2007, the European Parliament adopted a declara-
tion urging the Commission and the Council of Ministers to end the use of apes and wild-
caught monkeys and to establish a timeline to replace all non-human primates in scientific
experiments. 342 The Scientific Committee on Health and Environmental Risks (SCHER)
concluded that from a scientific point of view "developing and testing xenotransplantation
methodologies" was one of four areas in which "the use of non-human primates (NHPs), at
the present time, is essential for scientific progress". On 8 September 2010, the European
Parliament adopted a new Directive, in agreement with the Council, to revise Directive
86/609/EEC on the protection of animals used for scientific purposes (Directive 2010/63/
EU). It acknowledged that animals, including NHP, were still needed for scientific research.
On the other hand, the demand for human organs and tissues for transplantation far
exceeded the world supply. For every person who benefits from an organ transplant, there are
an estimated 5 to 10 other potential recipients for whom human organs are not available. 343
Xenotransplantation is viewed as a potential solution to the existing shortage of human
organs for transplantation.
Furthermore, with limited success in terms of patient survival or organ functionality,
experiments have been performed on transplants of bone marrow, hearts, neurons, cells 344
341. Taupitz J., M. Weschka (Edtrs) (2009), Chimbrids chimeras and hybrids in comparative Euro-
pean and International Research. Scientific, Ethical, Philosophical and Legal Aspects, Springer- Verlag
Berlin Heidelberg, pp. 98-100.
342. European Parliament (2007a): Declaration 0040/2007 on primates in scientific experiments. EP
reference number: DCL-oo40/2007/P6_TA-PROV(2007)00407, (http://ec.europa.eu/environment/
chemicals/lab_animals/ pdf/declaration_nhp_en. pdf).
343. Boneva R. S., T. M. Folks (2001), L. E. Chapman, "Infectious Disease Issues in Xenotransplanta-
tion", Clinical Microbiology Reviews, 14 (1): 1-14.
344. Sanal M. G. (2011), "Future of liver transplantation: non-human primates for patient-specific
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Raffaella Leoci
and other tissues from baboons, chimpanzees and pigs. However, pig and cow heart valves
have been commonly used in human patients. 345 Some heart patients owe their lives to valves
from pig hearts.
Pigs are a promising donor species because their organs are similar in size to humans,
they have large litters, grow quickly, and are far enough from humans on the evolutionary
tree to minimize infection risks. They are socially since many in our society regularly con-
sume them for food.
Despite several potential advantages over allotransplantation, xenotransplantation en-
compasses a number of problems. It is a common misconception that successful xenotrans-
plants involve animals that are most like humans, i.e. chimpanzees. In fact, chimps are not
so much like humans; and they are endangered, expensive to raise, and grow too slowly to be
of use for the thousands needing transplants. For example, at Loma Linda University Medi-
cal Center in California, on October 26, 1984, a 2-week-old infant known as "Baby Fae"
received a baboon heart transplant. She died after 20 days.
The potential to introduce infections across species barriers is another concern. Chimps
are also so unlike humans that the risks of transplanting an infectious virus are too high. 346
The human immune system reacts violently to pig organs and pigs contain ubiquitous
retroviruses that may adversely affect humans. Many bio tech companies are working to re-
solve these and other ethics issues that contribute to the xenotransplantation debate.
4.10 Intestines
The processing of animal intestines includes the following principal operations: separa-
tion of the intestinal complex into parts; removal of the intestinal contents and thorough
washing of the intestines ; fat extraction (skimming) ; and removal of the mucous membrane
(stripping) and other superfluous layers. The processed intestines are preserved with salt or
by curing. Slaughterhouses that do not process animal intestines preserve incompletely pro-
cessed (not cleaned and washed ) intestines by corning ( salting or soaking in brine). Salted
intestinal finished products are packed in wooden barrels, and dried intestinal finished prod-
ucts are packed in wooden boxes. Intestines may be utilized for: sausage casing, strings (for
suturing material, surgical ligatures, musical instruments, and racquets), and for use in the
manufacturing of heparin, and of intestinal mucus (SIS materials) .-
Sausage casings
Animal casings can be used for a variety of different purposes, such as surgical sutures,
collagen sheets (used for burn dressing), strings for musical instruments, casings, human
food, pet food, meat meal, tallow, and fertilizer.
organs from induced pluripotent stem cells", World Journal of Gastroenterology, 17 (32): 3684-90.
345. Ekser B., P. Rigotti, B. Gridelli, D. K. Cooper (2009), "Xenotransplantation of solid organs in the
pig-to-primate model", Transplant Immunology, 21 (2): 87-92.
346. Yoo D., A. Giulivi (2000), "Xenotransplantation and the potential risk of xenogeneic transmis-
sion of porcine viruses", Canadian Journal of Veterinary Research, 64 (4): 193-203; Boneva R. S., T.
M. Folks, L. E. Chapman (2001), Ibid., 14 (1): 1-14; Aynaud J. M. (2000), "Risks of infection in
xenotransplantation: what are they and how are they to be controlled?", Pathologie Biologic (Paris), 48
(4): 387-8; Meng X. J. (2003), "Swine hepatitis E virus: cross-species infection and risk in xenotrans-
plantation", Current Topics in Microbiology and Immunology, 278: 185-216.
92
Animal by-products (ABPs): origins, uses, and European regulations
Certainly the area having the highest economic value combined with volume of utiliza-
tion would be sausage casings. For these purposes sausage casings must be processed in a
specific way. 347 Some of them are used as container for the sausage mix and peeled off before
consumption while others are eaten with the sausage.
Both natural and artificial casings are soft cylindrical tubes used to contain sausage
mixes. Natural casings are obtained from animal intestines 348 derived from slaughtering.
Areas of the animal used to manufacture casings are the small and large intestine, gullet and
throat, urinary bladder, stomach, and rectum from most meat-producing animals. Many
factors influence the quality of the casing, such as health of the animal, specie, age of the
animal, fodder consumed, breed, conditions under which the animal was raised, portion of
the intestinal tract utilized, and how the product is handled and processed after the animal
is slaughtered. Casings are mainly derived from the small and large intestines of sheep, goats
and pigs, but also from cattle and horses. Small caliber natural casings are derived from the
small intestines of sheep, goats and pigs. They are processed in a way that makes them tender
and edible and are usually eaten with the sausage. The small intestines are detached from
adhering mesenteric tissue. The intestinal content is removed manually and the empty cas-
ings is flushed with water and subsequently de-slimed by using either manual or electrically
operated casing-cleaning machines. 349
Many other parts of the intestinal tract of slaughter animals can also be used for natural
casings but they are processed differently due to stronger and tougher casing walls. Because
of the hardness and toughness, those parties are generally considered unfit for human con-
sumption and are usually peeled off before consuming the sausages.
Only the small intestines of the gastro-intestinal tract of sheep and goats are normally
processed to be used as casings for many types of products (e. g. fresh frying sausages, frank-
furters, BBQ sausages, hot dogs and thin dried fermented sausages). These casings are pro-
cessed in a way that they become tender enough to be easily chewed.
Regarding pig intestines, several parts are processed into casings, and the most im-
portant is the small intestines. The large intestines having an average length of 3 m (pig
middles) and the cap are used as casings for coarse liver sausage and sometimes for salamis.
The "bung", the last part of the gastro-intestinal (average length of 0.8 m), is notable for
its strength and shape and is used as casing for products such as cervelat 350 and fine emul-
sified liver paste. The pig bladder can also be used for products such as black pudding or
gelatinous meat mixes.
347. Among others see: Gerrard F. (1969), Small goods production: a practical handbook on the manu-
facture of sausages and other meat products, Leonard Hill Books, London; Gayler P. (2011), Sausages,
Jacqui Small, London.
348. Anatomically the walls of the intestinal tract of slaughter animals consist of four layers of intesti-
nal tissue. These layers from inside to outside are: Mucose membrane (I), submucose membrane (II),
muscular layer (circular and longitudinal) (III) and serose membrane (IV). For natural casing manufac-
ture, one or more of these layers are removed during casing processing depending on the type of casing
(thin/ thick, edible/non-edible) to be fabricated.
349. The small intestines are passed through a set of rollers to loosen the tissue layers and to remove
the "slime" that is the internal layer of the intestine, basically the internal mucose or membrane. In the
slaughtered animal this membrane disintegrates rapidly and can easily be removed.
350. Cervelat, also cervelas, servelat or zervelat is a kind of finely chopped dry fermented sausage pro-
duced mainly in Switzerland, Alsace and in parts of Germany.
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As regards cattle, many parts of their gastro-intestinal tract are used as casings in sausage
production: "rounds" (small intestines) 351 are used for stuffing sausages such as lyoner 352 ,
liver and blood sausages and dried fermented beef products: the "middles" 353 are used for
dried fermented and precooked-cooked sausages such as "hunter's sausage" and coarse liver
sausage; the "blind gut" is also used for precooked-cooked sausages and raw-cooked prod-
ucts such as large bologna, etc. Beef bladders are used for "mortadella" and other specialties.
These natural casings are usually not eaten owing to their tough casing walls, although they
are edible.
Artificial casings are made of cellulose, collagen, and synthetic materials. They were
developed in response to the growing meat industries at the beginning of the 20th century.
At that time, in some countries, the supply of natural casings could no longer cope with the
demand for such natural casings. Artificial casings showed several advantages compared to
natural casing, as for example the negligible microbial contamination unnecessary refrigera-
tion, and absence of spoilage problems during transport and storage. Nowadays, for wide
sausage calibers, artificial casings are the material of choice, while for smaller caliber prod-
ucts, artificial and natural casings remain equally important.
According to their structure and composition of material, artificial casings can be sub-
divided into
1) casings made of natural materials, with two groups:
a) casings made of organic plant material, namely cellulose;
b) casings made of animal by-products, namely collagen.
2) casings made of synthetic substances deriving from thermoplastic materials which
can be subdivided into "polymer casings" and "plastic casings".
Collagen casings
This type of casings is fabricated from collagen, 354 which is obtained from the corium
layer of selected split cattle hides. The advantages are their standard diameter and strength
and that they can be "shirred", i.e. folded together, in long lengths and used for manual or
automatic filling stations without pre-soaking in water.
Synthetic casings
Synthetic casings are made of synthetic thermoplastic materials: Polyamide (PA), Poly-
ethylene (PE), Polypropylene (PP), Polyvinylidenchloride (PVDC) and Polyester (PET).
They can be manufactured with tailor-made properties but cannot be used for products
which have to undergo drying, ripening and fermentation, such as with dry sausages, since
they are impermeable for gases and water vapor.
351. Rounds are 40 m long and are normally readily available where cattle are slaughtered.
352. Lyoner or sausage is a sausage without lining. The recipe originally came from the French city of
Lyon - there she is Cervelas called.
353. The beef middles are separated from the "ruffle" (mesenteric fat), flushed out with water, trimmed
free of fat, turned inside out, slimed and salted. Beef middles include the "straight" casing (long, not
curved part) and are packed in sets each measuring about 17m after salting and composed of 5 pieces.
354. Collagen is an animal tissue fit for human consumption, the thin collagen casings are easy to chew
and "edible". They are an alternative to replace natural sheep, goat or thin pig casings.
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Animal by-products (ABPs): origins, uses, and European regulations
Glycosaminoglycans
These are aminopolysaccharides found in various forms throughout most animal tis-
sues. Heparin 355 is obtained from intestinal mucosa, mainly from sheep and pigs. It is used
commercially as a blood anticoagulant to prevent blood from clotting during operations
and during blood dialysis. Derivatives of heparin and similar glycoaminoglycans are used to
improve peripheral blood circulation. As a cosmetic heparin (applied topically) is used to
reduce bruising and sometimes as a moisturizer.
Suturing material
Catgut has been used by humans for centuries. It is extremely durable and strong, 356 and
can be used to suture, to string musical instruments, and to string tennis rackets.
After surgical operation, sutures hold the basic structural elements in their required sites
and provide necessary strength. They are characterized as biodegradable or non-biodegrad-
able. The first are used mainly for internal wound closures. The latter are used for closed
exposed wounds and are removed when the wound is quite healed. Sutures may be of a
natural or synthetic variety.
A catgut 357 suture is a suturing material made from the natural fiber found in the walls
of animal intestines that is naturally degraded by the body's own proteolytic enzymes. It is a
popular material because it is absorptive and can be sterilized. Historical references to the use
of twisted intestinal materials for surgical suture go back to the 2nd century A.D. and its use
in musical instruments far longer. The name catgut and its use as suture first appeared in the
writings of Claudius Galenus around 50 AD. Sheep have historically been used as a source
for catgut, although dogs, cattle, goats, hogs, horses, mules, or donkeys have been used as
well. Absorbable sutures can be plain catgut, which usually holds its strength for about 10
days or catgut treated with chromic acid (chromic sutures) that has a slower absorption and
keeps its strength for 20 days. Catgut surgical sutures, both chromic and plain, can be used
in general closure, ophthalmic, orthopedics, dentistry, gynecology and gastro surgeries.
355. Heparin is a polymer classified as a mucopolysaccharide or a glycosoaminoglycan. It is biosyn-
thesized and stored in mast cells of various mammalian tissues, particularly liver, lung and mucosa.
Commercial heparin is chiefly isolated from beef lung or pork intestinal mucosa. See: Linhardt R. J.,
N. S. Gunay (1999), Production and chemical processing of low molecular weight Heparins, Seminar in
Thrombosis and Hemostasis, Thieme Medical Publishers, 25 (3), N.Y.
356. Catgut strings are made preferably from the small intestines of sheep. To produce the strings, the
guts are cleaned and cut into about 15 m strands before being chemically treated to preserve them.
Each string is made up of 15 individual strands which are spun very tightly together. Then they are
dried out in a humid room to prevent cracking. It is a painstaking process that takes six weeks from
start to finish.
357. Catgut sutures, despite the name, perhaps have nothing to do with cats. The word appears to have
come into common use several hundred years ago. Although it would be possible to derive catgut from
cats, the yield would not be impressive when compared to that of a larger animal. The origins of the
term "catgut" appear to come from "kitgut," a word derived from "kit" for "fiddle." However, since a
small cat is also called a Kit, some say this was the evolutionary pathway from "Kat gut" to "Catgut".
Kitgut fiddle strings were made from the processed intestines of animals such as sheep, and over time,
the word was corrupted into "catgut." Another maintains that the best lamb gut musical strings came
from Catagniny, Germany. Top musicians demanded Catagniny Gut, which eventually evolved into
"Catgut". Confusion about the term persists to this day.
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Raffaella Leoci
Catgut, however, can possibly lose its tensile strength, have tainted purity, and be
expensive. 358 Collagen was invented to overcome the disadvantages of catgut. The flexor
tendons of beef are converted into dispersed fibrils that are extruded and reconstituted to
form collagen sutures.
There is debate about whether to continue using catgut in a medical setting, since cot-
ton is usually cheaper and wounds closed with either cotton or synthetic threads (as polyg-
lycolic acid and polyglactin 359 ) are less prone to infection. Biosecurity Australia, an agency
within the Australian Department of Agriculture Fisheries and Forestry, identifies catgut
as a "high TSE risk product" since it is derived from bovine or ovine intestinal tissue. 360 In
Europe and Japan, gut sutures have been banned since 2002 due to concerns that they could
transmit bovine spongiform encephalopathy (mad-cow disease), even though the herds from
which gut is harvested are certified BSE-free. In Italy the Decree of 16 July, 2002 from the
Ministry of Health 361 prohibited "the use, importation and placing on the Italian market of
medical devices, in the form of absorbable surgical sutures (catgut), obtained from bovine,
sheep and goats intestine, as well as the use, importation and placing on the Italian market
of medical devices obtained from materials of bovine origin coming from the dura mater,
brain and spinal cord".
The use of animal intestines to make thread for sewing footwear and fur clothing has
been known from very ancient times.
Musical string
For thousands of years, the choices of musical string materials were few. Usually it was
limited to some indigenous material that was suitable to the task. For a long period, catgut
was the most common material for harps, lutes, violins, and violas strings, as well as for the
surface of older marching snare drums.
No one knows exactly when catgut was first used for musical strings. According to leg-
end, Apollo was the first string maker. One day, when he came across a tortoise, he was in-
spired to make the first lyre, and he used the poor animal's own intestines for the strings. The
first evidence of gut string use dates back to 1 823. The architect James Burton Jr. purchased
some musical instruments 362 in Thebes (Greece) for his early musical instrument collection.
358. Doctors at the New Orleans Charity Hospital had found that wounds stitched together with
"ordinary cotton thread" were less likely to become infected than those sutured with catgut or silk. An-
other advantage: cotton is not absorbed and will hold when a wound takes a long time to heal — catgut
may disappear in a little over a week.
359. Leroux N., E. Bujold (2006), "Impact of chromic catgut versus polyglactin 910 versus fast-
absorbing polyglactin 910 sutures for perineal repair: a randomized, controlled trial", American Journal
of Obstetrics and Gynecology, 194 (6): 1585-90; Kettle C, R. B. Johanson (2010) , Absorbable synthetic
versus catgut suture material for perineal repair, in The Cochrane Library, Issue 2,. Chichester, UK: John
Wiley & Sons, Ltd. Search date 1999.
360. Department of Agriculture Fisheries and Forestry Australia, Measures to address additional TSE
concerns with veterinary vaccines and other high risk biologicals, November 2001.
361. Ministero della Salute, "Misure di protezione nei confronti delle encefalopatie spongiformi
trasmissibili relativamente ai dispositivi medici" ("Protective measures with regard to transmissible
spongiform encephalopathies in relation to medical devices", Official Gazette of the Italian Republic
n. 192 of 17 August, 2002).
362. Some are currently in the British Museum at London.
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Animal by-products (ABPs): origins, uses, and European regulations
These interesting relics had been found in a tomb at Thebes. One stringed instrument, when
found, "was nearly perfect, and had parts of the catgut of all the four string". 363 An Egyp-
tian harp had gut strings that still made a tone after some two thousand years in storage. 364
Most musical instruments produced today use strings with cores made of other materials,
generally steel or synthetic polymer. Catgut strings are still commonly preferred in concert-
tension pedal/grand and some lever harps, because they produce a richer, darker sound as
well as withstand high tension within low alto, tenor, and high-bass ranges.
Strings for tennis racquet
On the market there are many different types of strings; selecting a string can be chal-
lenging as there are hundreds of strings to choose from. Making gut strings from cows re-
quires a complex process. Because of this, gut is the most expensive string on the market. 365
Gut string is very popular among professional players because of its elasticity, tension stabil-
ity, and liveliness. Because of its high price, however, gut is not recommended for the average
recreational player. It is also not very durable and is very sensitive to moisture.
Synthetic gut strings provide different characteristics such as durability, spin, feel, pow-
er, etc. They can be made from nylon, polyester and Kevlar, and multifilaments, etc.
In recent years hybrid strings are gaining popularity as more players are looking for a
blend of string qualities. By selecting different hybrid combinations of string, players can
fine-tune the playability, comfort, durability, liveliness, and control offered by the stringbed.
For instance, heavy hitting players can find a good combination of durability and playability
with a polyester main string and natural gut or premium synthetic cross string hybrid.
Small intestine submucosa
The mucous membrane is the weakest of the four layers that make up the intestines, and
it is removed when processing the raw material. The submucous layer is the strongest and it
is always left intact. 366 The muscular and serous layers are removed or left depending upon
how tough they are (which is determined by the section of the intestine) and the purpose
of the finished product. Intestines are usually washed in the slaughterhouse. The intestine
wash water contains valuable mucus that can be recovered, cleaned for pathogen material,
concentrated to 150 centipoises, and spray dried.
Porcine small intestine submucosa (SIS) 367 can be implanted into humans to help repair
363. Limbird J. (1836) (Publisher), The Mirror of Literature, Amusement, and Instruction, Vol. XXVIII,
London, p. 98 (Egyptian Antiquities: Musical Instruments).
364. Wilkinson J. G. (1841), A second series of the Manners and customs of the ancient Egyptians:
including their religion, agriculture, & c. Derived from a comparison of the paintings, sculptures, and
monuments still existing, with the accounts of ancient authors, Vol. I, J. Murray, London.
365. It seems that it takes about four cow's guts to string an average racquet.
366. Bejjani G. K., J. Zabramski (2007), "Safety and efficacy of the porcine small intestinal submucosa
dural substitute: results of a prospective multicenter study and literature review", Journal of Neuro-
surgery, 106 (6): 1028-33; Badylak S., K. Kokini, B. Tullius, A. Simmons-Byrd, R. Morff (2002),
"Morphologic study of small intestinal submucosa as a body wall repair device", Journal of Surgical
Research, 103 (2): 190-202.
367. Zheng M. H., J. Chen, Y. Kirilak, C. Willers, J. Xu, D. Wood (2005), "Porcine small intestine
submucosa (SIS) is not an acellular collagenous matrix and contains porcine DNA: Possible implica-
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Raffaella Leoci
everything from eyelids to eardrums. The SIS material, which is not rejected by the human
and animal body, signals the patient's own cells to begin to grow and repair tissues. There
have been studies regarding this phenomena.
Sandusky Jr. and others 368 implanted a small caliber vascular graft from porcine small
intestine submucosa in a canine carotid artery. They later compared it with an autogenous
saphenous vein graft that was implanted in the contralateral carotid artery. The results of
these studious indicate that the SIS graft is similar to saphenous vein graft in the dog.
Badylak and others 36 ' tested the use of autogenous small intestinal submucosa (SIS) as a
large diameter (10 mm) vascular graft in the infrarenal aorta of 12 dogs. They conclude that
autogenous small intestinal submucosa can be successfully used as a large diameter arterial
graft in the dog and is worthy of further investigation.
Kropp and others 370 determined the feasibility of promoting urinary bladder regenera-
tion with porcine-derived small intestinal submucosa (SIS). This study further supports the
concept of bladder regeneration and suggests that SIS may be a viable material for bladder
augmentations.
Alvin B. Rutner and others 371 described their surgical technique and results with implanta-
tion of processed porcine small intestine submucosa (SIS) as a pubo-vaginal sling in 1 52 consecu-
tive female patients with stress urinary incontinence (SUI). They concluded that processed SIS is
strong, durable, biocompatible, infection resistant, and gradually replaced by host tissues.
However, there are doubts about the use of SIS on people. 372 The long-term efficacy of
pubo-vaginal sling (PVS) procedure with porcine small intestinal submucosa (SIS) implant
was retrospectively assessed by Siracusano and others. 373 They concluded that PVS procedure
using SIS cannot offer a durable option for the treatment of SUI as reported by the current
mini-invasive techniques.
tions in human implantation", Journal of Biomedical Materials Research Part B: Applied Biomaterials,
73B (1): 61-67; Dejardin L. M., S. P. Arnoczky, B. J. Ewers, R. C. Haut, R. B. Clarke (2001), Tissue-
Engineered Rotator Cuff Tendon Using Porcine Small Intestine Submucosa. Histologic and Mechanical
Evaluation in Dogs, Presented at the 26th annual meeting of the AOSSM, Sun Valley, Idaho, June
2000 (in The American Journal of Sports Medicine, 2001, vol. 29, pp. 175-184); McDevitt C. A., G. M.
Wildey, R. M. Cutrone (2003), "Transforming growth factor-(31 in a sterilized tissue derived from the
pig small intestine submucosa", Journal of Biomedical Materials Research Part A, vol. 67A (2) : 637—640.
368. Sandusky Jr. G. E., S. F. Badylak, R. J. MorfF, W. D. Johnson, G. Lantz (1992), "Histologic
Findings After In Vivo Placement of Small Intestine Submucosal Vascular Grafts and Saphenous Vein
Grafts in the Carotid Artery in Dogs", American Journal of Pathology, 140 (2): 317-324.
369. Badylak S. E, G. C. Lantz, A. Coffey, L. A. Geddes (2002), "Morphologic study of small intesti-
nal submucosa as a body wall repair device' ', Journal of Surgical Research, 103 (2): 190—202.
370. Kropp B. P., B. L. Eppley, C. D. Prevela, M. K. Rippy, R. C. Harruff, S. F. Badylak, M. C. Adams,
R. C. Rink, M. A. Keating (1995), "Experimental assessment of small intestinal submucosa as a blad-
der wall substitute", Urology, 46 (3): 396-400.
371. Rutner A. B., S. R. Levine, J. F. Schmaelzle (2003), "Processed porcine small intestine submucosa
as a graft material for pubovaginal slings: durability and results", Urology, 62 (5): 805—809.
372. Schaefer M., A. Kaiser, M. Stehr, H. J. Beyer (2013), "Bladder augmentation with small intestinal
submucosa leads to unsatisfactory long-term results", Journal of Pediatric Urology, published online 17
January 2013.
373. Siracusano S., S. Ciciliato, N. Lampropoulou, A. Cucchi, F. Visalli, R. Talamini (201 1), "Porcine
small intestinal submucosa implant in pubovaginal sling procedure on 48 consecutive patients: long-
term results", European Journal of Obstetrics & Gynecology and Reproductive Biology, 158 (2): 350-353.
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Animal by-products (ABPs): origins, uses, and European regulations
4.11 Lungs
Pig, calf and lamb lungs, as foods, are mainly used to make stuffing and some types of
sausages and processed meats.
In 1963, the first single human lung transplant was performed at the University of
Mississippi, 374 but the patient died within days because of renal failure and general debility.
Others transplants followed with increasing success. 375
Heparin can also be extracted from the lungs for use as an anti-coagulant, gangrene pre-
ventative, and as meal (as already mentioned above). Heparine is also prepared from bovine
lung and bleached during processing. It has been traditionally used as an anticoagulant and
binds to antithrombin III, a naturally occurring plasma protease inhibitor, accelerating the
rate at which antithrombin III inhibits coagulation proteases. 376
Aprotinin is another compound obtained from bovine lungs and available on the mar-
ket as lyophilized powder. The term is typically used when describing the protein derived
from bovine lung.
Aprotinin was isolated independently from two laboratories and originally named Bo-
vine Pancreatic Trypsin Inhibitor and Trypsin-kallikrein Inhibitor. It is a protein consisting
of 58 amino acids, arranged in a single polypeptide chain that is crosslinked by three di-
sulfide bridges. It is an antifibrinolytic medicine, which prevents excessive blood loss. Apro-
tinin is a competitive serine protease inhibitor that inhibits trypsin, chymotrypsin, kallikrein
and plasmin. 377
Aprotinin is producted by lungs. The manufacture involves extraction from minced
tissue, separation of solids, clarification, ultrafiltration and concentration and fractionation
by reverse-phase chromatography. Extration, removal of solids, clarification by alluvial filtra-
tion, and ultrafiltration are standards tasks. Following concentration, the final yield volume
is about 10 liters from 100 kg of lung.
4.12 Meat and bone meal
Meat and bone meal (MBM) revenue is an important aspect to the profitability of ren-
dering operations and the meat industry in general. Usually destined for disposal, bio-oils
and higher value chemical products can be extracted from meat and bone char for use as
internal energy or for sale.
374. HardyJ. D. (1999), "The First Lung Transplant in Man (1963) and the First Heart Transplant in
Man (1964)", Transplantation Proceedings, 31: 25—29.
375. Arcasoy S. M., R. M. KotlofF (1999), "Lung Transplantation", New England Journal of Medicine,
340 (14): 1081-1091.
376. Melo F. R., M. S. Pereira, D. Foguel, P. A. S. Mourao (2004), "Antithrombin-mediated Anticoag-
ulant Activity of Sulfated Polysaccharides. Different Mechanisms for Heparin and Sulfated Galactans",
The Journal of Biological Chemistry, 279 (20): 20824-20835; Bjorkl., U. Lindahl (1982), "Mechanism
of the anticoagulant action of heparin", Molecular Cellular Biochemistry, 48 (3): 161-182; Jordan R., D.
Beeler, R. Rosenberg (1979), "Fractionation of low molecular weight heparin species and their interac-
tion with antithrombin", The Journal of Biological Chemistry, 254 (8): 2902-2913.
377. Kassell B., M. Laskowski Sr. (1965), "The basic Trypsin inhibitor of bovine pancreas: V. The
disulfide linkages", Biochemical and Biophysical Research Communications, 20 (4): 463-468; Kassell B.
(1970), "Bovine Trypsin-Kallikrein inhibitor (Kunitz Inhibitor, Basic Pancreatic Trypsin Inhibitor,
Polyvalent Inhibitor from Bovine Organs)", Methods in Enzymology, 19: 844-852.
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Raffaella Leoci
The most mature and suitable technologies for implementation within the meat pro-
cessing industry are Pyrolisis, 378 Fluidized bed reactor, Anaerobic digestion, and Co-firing/
Incineration.
Pyrolisis
Pyrolisis is a thermochemical decomposition of organic material at various tempera-
tures (high, low, or medium) in the absence of oxygen. The process is very widespread in
the chemical industry: for example, to produce charcoal, activated carbon, methanol, and
other chemicals from wood; to convert ethylene dichloride into vinyl chloride to make PVC;
to produce coke from coal; to convert biomass into syngas and biochar; to turn waste into
safely disposable substances; and for transforming medium-weight hydrocarbons from oil
into lighter ones like gasoline.
Pyrolysis typically occurs under pressure and at operating temperatures above 430 °C
when organic materials are transformed into gases, small quantities of liquid, and a solid
residue containing carbon and ash.
The main potential of this technology is the production of a liquid fuel (bio-oil) suit-
able for transport and storage. Its advantage of over other methods of energy extraction
from MBM is the milder operating conditions, around 500°C instead of 800-900°C for
gasification, and the very short processing times compared to the several weeks required for
anaerobic digestion. Key to commercial success seems to be the extraction of higher value
chemical by-products that occur naturally during the pyrolysis of biomass, in addition to
the bio-oil itself.
MBM typically is a fine powder and has low moisture content (about 5%), making it
a good feed for pyrolysis. But a basic financial analysis indicates this use of MBM is not yet
viable and the process will not be considered while a market for MBM as a animal feed food
ingredient remains.
Fluidized bed reactor
Another suitable technology for implementation within the meat processing industry
is the fluidized bed reactor. Unfortunately, the small feed size requirements to aid reaction
rate may be a problem considering of the energy required for particle size reduction, with
the exception of MBM.
Anaerobic digestion
Anaerobic digestion of organic material produces methane and carbon dioxide gases
and fertilizer. This process does not deactivate pathogens since the maximum temperature
attained in commercial composting is below that required for pathogen inactivation. Conse-
quently it is possible that the presence of "high risk" materials (i.e., brain, spinal cord, etc.)
may not be allowed to enter the process stream. MBM may require prior heat treatment
(pasteurization) in order to meet further use regulations, and this involves significant ad-
ditional costs to the anaerobic digestion process.
378. McGlashan S. A. (1997), Industrial and Energy Uses of Animal By-Products, Past and Future, En-
vironment and Co-Products Meat and Livestock Australia, Ltd. (http://assets.nationalrenderers.org/
essential_rendering_ industrial_and _energy.pdf).
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Animal by-products (ABPs): origins, uses, and European regulations
Co-firing/Incineration
Co-firing/Incineration can be used in the production of cement where co-firing/incinera-
tion of MBM offers several advantages over other disposal options. This substitution not only
provides a method of energy recovery but also reduces net greenhouse gas emissions by replac-
ing coal with a "carbon neutral" fuel. Derived from a biomass, is considered carbon neutral
because the carbon released upon combustion was the same absorbed from the atmosphere
during the growth of the organism. Another advantage is that the resultant ash is incorporated
in the final cement product, which reduces the amount of solid waste ending up in landfills.
MBM inclusion in concrete and asphalt construction composites appear to have some
promise. Further study is needed using MBM in construction applications.
4.13 Ovaries
There are many organs and glands in the body of animals: thyroid, adrenal, thymus,
testis, ovary glandular, etc. Glands contain hormones, amino acids, vitamins, enzymes, min-
erals, neurotransmitters, and many others compounds.
Past and modern ealers have used tissue extracts, in addition to herbs and plants, in the
fight against disease. For example, desiccated thyroid is still used by many alternative prac-
titioners in the management of hypothyroidism. Many people consume glands, perceiving
them to be a source for natural hormones, vitamins, etc. For example, they think that eating
brain tissue could improve mental function and eating heart tissue improves heart function,
etc. It is known that Thyroid glandular could improve thyroid function in those who are
hypothyroid. But one cannot always apply this principle to other glands.
There are no published studies about the safety or effectiveness of bovine ovary extract
in humans. Many think that bovine ovary extract stimulates the pituitary gland resulting in
an increase in prolactin and growth hormone levels.
Ovary extracts have various activities: ovary extracts may influence estrogen levels. Prop-
erties of ovarian extracts have been studied since before World War II. 379 The ovary is made
up of many distinct parts: the corpora lutea, the maturing graafian follicles, the interstitial
tissue, and the primary, or immature follicles. Progesterone has been obtained from the cor-
pora lutea. The corpora lutea also yields a substance with estrogenic power that is thought to
be estrone. Progesterone and estrogen can be extracted from pig ovaries. They may be used
to treat reproductive problems in women.
Relaxin, a substance with specific action on pelvic ligaments, is taken from the ovaries of
pregnant sows and is often used during childbirth. The effects of ovarian extracts containing
relaxin on various types of smooth muscle have been investigated by Miller and others. 380
Administration of the extracts to dogs produced hypotension, bradycardia, inhibition of
ureteral motility, and tachyphylaxis.
Many researchers have studied the properties of ovarian extracts. Let us remember some
of them. According to Spanel-Borowski 381 ovarian extracts appear to exert influence on
379. Marlow H. W. (1939), "Properties of ovarian extracts", Endocrinology, 25 (5): 793-797.
380. Miller J. W, A. Kisley, W. J. Murray (1957), "The Effects of Relaxin-Containing Ovarian Extracts on
Various Types of Smooth Muscle", Journal of Pharmacology and Experimental Therapeutics, 120 (4): 426-437.
381. Spanel-Borowski K., S. Weis (2007), "Ovarian Extract have an effect on Chemotaxis of Blood
Granulocytes. A Study Conducted in Superovulated Golden Hamsters", Reproduction in Domestic Ani-
mals, 25 (5): 269-276.
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chemotactic activity and adherence of blood granulocytes. Heilbrunn and others 382 pointed
out the antimitotic and carcinostatic action of ovarian extracts.
4.14 Proteins
For non-food proteins there are many potential applications. Among these we remem-
ber the following:
• Protein hydrolysis products
• Proteins for Plastics
• Protein-based adhesives
• Protein surfactant
a) Protein hydrolysis products
Proteins are made up of chains of amino acids linked by peptide bonds and folded in
a variety of complex structures. Protein hydrolysis is the breakdown of protein into smaller
peptides and free amino acids. Protein hydrolysates, also called peptones, are the result of the
hydrolysis process on protein material.
Peptones are obtained from enzyme catalysed hydrolysis and hydrolysates from acid
catalysed hydrolysis. Meat peptone is used in biotechnology as a nutrient in vaccine pro-
duction and in microbiological culture medium. Casein peptone is also used as a culture
medium while casein hydrolysates are used as an amino acid supplement (a nutriceutical)
and in cosmetics.
As starting material, commonly used protein sources, include meat, casein and whey
(milk proteins), gelatin, and soy beans. Hydrolysis of the starting material is accomplished
by utilizing acid 383 or enzymatic 384 or microbial 385 hydrolysis.
b) Proteins for plastics
Demand of environmentally friendly plastics manufactured from biodegradable materi-
als is growing continuously. 386 Plastic films can be made from thermoplastic polymers by
blowing or casting or molding. Extrusion is used for collagen products: a purified and acidi-
fied aqueous suspension is extruded into a coagulating bath. Thermoplastic extrusion is not
employed for protein-based films. A cast film is made by laying a solution of the polymer
382. Heilbrunn L. V, W. L. Wilson, T. R. Tosteson, E. Davidson, R. J. Rutman (1957), "Hie Antimi-
totic and Carcinostatic Action of Ovarian Extracts", Biological Bulletin, 113 (1): 129-134.
383. Adler-Nissen J. (1979), "Determination of the degree of hydrolysis of food protein hydrolysates
by trinitrobenzenesulfonic acid", Journal of 'Agricultural and Food Chemistry, 27 (6): 1256-1262.
384. Adler-Nissen J. (1976), "Enzymatic hydrolysis of proteins for increased solubility", Journal of
Agricultural and Food Chemistry, 24 (6): 1090-1093; Adler-Nissen J. (1986), Enzymatic hydrolysis of
food proteins, London: Elsevier Applied Science Publishing.
385. Ferrero M. A. (2000), Protein Hydrolysis: Isolation and Characterization of Microbial Proteases, in
John E Spencer and Alicia L. Ragout Spencer (Edts.), Food Microbiology Protocols, Series: Methods in
Biotechnology, Vol. 14, Springer Science+Business Media, Secaucus, N. J. (USA): 227-232.
386. Kolybaba M., L.G. Tabil, S. Panigrahi, W. J. Crerar, T. Powell, B. Wang (003), Biodegradable
Polymers: Past, Present, and Future, presented at the 2003 CSAE/ASAE, Annual Intersectional Meet-
ing, Sponsored by the Red River Section of ASAE, Fargo, North Dakota, USA, October 3-4, 2003.
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Animal by-products (ABPs): origins, uses, and European regulations
dissolved in a volatile solvent on a flat surface. Solvent evaporation leaves a polymer film.
Forming the polymer relies on a solvent casting that uses water, acidic water, alkaline water,
or aqueous ethanol as the solvent (depending on the type of protein).
Currently, the most mature technologies use wheat and cornstarches, soy proteins, and oil-
derived esters as raw material. A wide range of proteins have been used to produce edible and/
or biodegradable casings and coating for food, pharmaceuticals, and industrial products. For a
long time protein films have been used as meat casing. Other uses include collagen casings, gel-
atin capsules, and microcapsules. Lately, the search for biodegradable polymers for packaging
and food wraps has resulted in the growth of research and in a development of activity. 387 How-
ever, few commercially produced biodegradable plastics are price competitive with traditional
oil-derived plastics such as polyethylene and polystyrene. Moreover, bio-based plastic derived
from fermentation processes, i.e. as for protein-based plastics, are generally more expensive
than those manufactured via chemical processes. Furthermore, most biodegradable plastics
are mechanically inferior to polyethylene and polystyrene, though the properties of protein
films can also be modified by cross-linking the protein molecules and modifying the molecular
structure using various physical and chemical processes. The hydrophilic nature of protein
films implies that they have poor moisture barrier properties, though structural modification
or the addition of lipids, waxes and so on can decrease the water vapor transmission rate. 388
Protein films tend to be brittle, so some plasticizers can be used, such as propylene
glycol, glycerol, triethylene glycol, sorbitol, and sucrose, etc. The use of plasticizers tends to
decrease film stiffness and tensile strength, while increasing elasticity and permeability.
c) Proteins as adhesives
Protein streams from rendered co-products are well suited to adhesive applications due to the
large number of available chemical functionalities associated with the amino acids that are useful
for bond formulation. The functional group can then form both physical bonds with the sub-
strate material or can form true chemical bonds with the functional group found on the substrate.
The use of waste protein as a raw material in the manufacture of adhesives for wood
composites has been the subject of extensive study in many countries over the past 50 years.
Animal protein-based adhesives can be derived from animal blood, although some involve
the use of specific proteins primarily selected from collagen and blood albumin.
Mechanisms for using proteins as adhesives are gel penetration (the gel penetrates the
pores of substrates), protein-substrate bonding, thermoset encapsulation and high- adhesion
protein structures.
Animal-based adhesives have been used since the early 1800s and consumption peaked
at about 70,000 tons in the USA in 1948. After World War II, low cost synthetic adhesives
quickly infiltrated the market replacing those products from animals that were because tech-
nically inferior and more expensive.
387. The state of the art for vegetable and animal protein film applications are reviewed by Gennadios
A. (2002) (Edt.), Protein-based films and coatings, CRC Press, Florida (USA).
388. Tharanathan R. N. (2003), "Biodegradable films and composite coatings: Past, present, and fu-
ture", Trends in Food Science and Technology, 14 (3): 71-78; Aminabhavi T. M., R. H. Balundgi, P. E.
Cassidy (1990), "Review on biodegradable plastics", Polymer Plastics Technology and Engineering, 29
(3): 235-262; Fomin V. A. (2001), "Biodegradable polymers, their present state and future prospects",
Progress in Rubber and Plastics Technology, 17 (3): 186-204.
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The largest adhesive market dominated by urea-formaldehyde 389 and phenol-formalde-
hyde 390 resins is for the production of wood products such as plywood. Consequently, the
primary target market is for protein-based adhesive formulations that may act as substitutes
for formaldehyde resins is the plywood market, but there are few large-scale uses of waste
animal proteins inhibiting development of this market. The costs of transforming waste
animal protein into a form that is suitable for use in adhesive formulations makes this use
economically attractive. But it is not just a matter of cost. There is also the problem of the
low water resistance of protein-based adhesives and resulting accelerated bio-deterioration of
the product. Research into cross-linking processes and reactive addition or modification of
functional groups may overcome some aspect of the adhesives poor water resistance.
d) Protein surfactants
"Surfactant" is a contraction of the term "surface active agent". This agent is a chemical
that is structurally attracted to the interface between two phases of matter. In other word,
surfactants are compounds that lower the surface tension of a liquid, the interfacial tension
between two liquids, or the surface tension between a liquid and a solid. 391 The surfactants
are characterized by their amphipathic 392 structures: one part of the molecule will be lypo-
phobic, while the other part will be lypophillic. Proteins, as already mentioned, are polymers
with a lot of amino acids that have a number of functional groups that can interact with two
sides of an interface or that can be derivatized with another compound to form an amphi-
pathic structure. 393
Surfactants are used as wetting agents, foaming and anti-foaming agents, emulsifiers,
and dispersion and aggregation agents.
Protein-based surfactants are used in the cosmetic and personal care industry. 394 At pre-
sent, vegetable derived proteins are almost always preferred in the personal care market. How-
ever, surfactant production is dominated by synthetic surfactants made from petroleum.
4.15 Stomach and tripe
Ruminants are herbivores that utilize, in their forestomach (rumen), a symbiotic re-
lationship with microorganisms in order to exploit fibrous feeds as a source of energy and
nutrients. 395
389. Urea-formaldehyde resins are used as adhesives for indoor wood composite products as they are
less water-resistant.
390. The phenol-formaldehyde resins are used for outdoor applications for their water resistance and
to minimize the effects of formaldehyde emission.
391. See: Rosen M. J., J. T. Kunjappu (2012), Surfactants and Interfacial Phenomena, 4 th Edition, John
Wiley & Sons, Hoboken, New Jersey.
392. Amphiphile from the Greek amphis (both) and philia (love, friendship). It is a term describing a
chemical compound possessing both hydrophilic and lipophilic properties. Such a compound is called
amphiphilic or amphipathic.
393. Krantz D. D., R. Kagan, S. Lawrence Zipursky (1991), "Amphipathic (3 Structure of a Leucine-
rich Repeat Peptid", The Journal of Biological Chemistry, 266 (25): 16801-16807.
394. Nnanna I. A., Jiding Xia (2001), Protein-Based Surfactants - Synthesis, Physicochemical Proper-
ties and Applications, CRC Press, Florida (USA).
395. Microbial fermentation of ingested plant materials is a crucial step in the digestion of feed by the
host animal. Most microorganisms have different roles in feed digestion and act synergistically to fer-
104
Animal by-products (ABPs): origins, uses, and European regulations
Ruminant stomachs have four compartments: rumen, reticulum, omasum and an abo-
masum that corresponds to the omnivore stomach. Stomachs are abundant as slaughterhouse
by-product and if they are not used as a waste material can create environmental pollution.
a) Ruminant stomach as food
Tripe is a generic term referring to the stomach of various ruminant animals especially
cattle, sheep, goats, and deer. Ruminant animals have multi-chambered stomachs 396 , and
tripe generally comes from either the rumen or reticulum chambers. In other word, tripe
is the culinary term for the stomach tissue, or offal, of most ruminant animals. The rumen
and reticulum are those most often used as food. They are generally processed at the place
of collection by washing, scalding and bleaching. They are suitable for poaching or braising,
and can be used in sausages and processed meat and sometimes can also be sewn to form a
casing and stuffed.
Tissues from the rumen are usually quite smooth, and are often known as "flat tripe."
The reticulum chamber produces the higher-priced honeycombed varieties, which are ap-
preciated for their texture and for their flavor.
Tissues taken from the omasum or reticulum are frequently known as "book" or "leaf"
tripe. Abomasums tissues are often referred to as "green" because they can contain undi-
gested food (grasses, leaves, or shrubbery that the animal ate shortly before it died can often
be found in this part of stomach). In the USA and Europe most processed abomasum tis-
sues are set aside for use in pet food, as they are a very cheap means of delivering protein to
domestic animals.
The health benefits and nutritive content of stomach tissues vary depending on the type
of animal and on the chamber from which they are derived. However, in most cases, the
stomach tissues are high in iron, calcium, and zinc and are very low in fat and calories, in
addition to being excellent sources of protein, vitamin B 12 .
A 100 g serving of tripe provides 1 1.8 g of protein, or 18,15% of the 65 g FDA daily
value; 4,2 g of fat, or 6% of the 65 g daily value; and 1.5 g saturated fat, or 7,1% of the 20
g daily value. A 100 g serving of tripe provides 81,1 mg calcium, or 8% of the 1,000 mg
FDA daily value. A 100 g serving of tripe provides 1.8 mg of zinc, or 11% of the 15 mg
FDA daily value. The selenium content in a 100 g of tripe is 1 1.9 meg, or 17 percent of the
70 meg daily value. 397
The United States Department of Agriculture (USDA) 398 recognizes two types of tripe:
honeycomb (Items No. 726 and 727) and other.
ment plant carbohydrates and proteins. Microbial populations change with feed type.
396. The cellulose and hemicellulose cannot be digested by mammalian enzymes and require bacteria,
protozoa and fungi in order to break down their. The rumen contains that microorganism and en-
zymes. The broken down material then moves to the omasum which acts as a filter and only enables
very small particles to move on to the abomasum while larger particles return to the rumen/reticulum
for further break down. The abomasum is the "true" stomach and acts like the monogastric stomach
by producing acid and protelytic enzymes.
397. Tyler Herbst S., R. Herbst (2007), The New Food Lover's Companion, Barron's Educational Series,
Inc., New York.
398. United States Department of Agriculture, Agricultural Marketing Service Livestock and Seed Pro-
gram, "Institutional Meat Purchase Specifications - For Variety Meats and Edible By-Products - Series
700", (1993), Washington, DC 20250.
105
Raffaella Leoci
Item No. 726 - Beef Tripe, Scalded, Bleached (Denuded) - The paunch with or with-
out the "honeycomb" reticulum will be scalded and washed absolutely free of any foreign
material and bleached with an FSIS approved bleaching solution. The color may range from
white to a light pale yellow. The dark internal lining will be removed.
Item No. 727 - Beef Tripe, Honeycomb, Bleached - The "honeycomb" reticulum will
be removed from the paunch by cutting along the seam connecting the two sections of the
stomach. The dark internal lining will be removed and the tripe will be scalded and bleached
to a creamy white color.
Europeans distinguish between four different types of beef tripe, and use all four.
• Plain Tripe or Rumen. Comes from the first stomach (called the "panes" in French)
and it is considered the least desirable tripe amongst tripe fans;
• Honeycomb Tripe or Reticulum (called "reseau" in French) comes from the lower
part of the second stomach. Pocket Tripe also comes from the second stomach;
• Book Tripe or Omasum (called "le feuillet" in French, "centopelli" or "fogliolo",
or "millefogli" or "bibbia" in Italian) comes from the third stomach;
• Reed Tripe or Abomasum (called "caillette" or "franche-mule" in French,
"lampredotto" 39 ' or "fisarmonica" or "frasame", or "riccia", or "frangiata", or "qua-
glio", or "riccioletta" in Italian). Comes from the fourth stomach. This is the stom-
ach that rennet is obtained from in calves. In Florence (Italy), they even distinguish
2 separate parts of the fourth stomach: the "spannocchia" and the "gala".
Pig stomachs are composed mainly of smooth muscle and collagenous connective tissue.
They are cleaned and scalded to remove the mucosa lining and so are suitable for braising,
and sometimes also used as a casing for sausages. They are processed in two ways. If the
stomachs are to be incorporated into meat mixes for sausage, they are scalded before further
processing, whilst, if they are used as casings, only a small opening is made, through which
they are cleaned by flushing with plenty of clean water. Pork tripe is called "trippetta" in Ital-
ian and is often considered to be cat food.
b) Rumen and rennet
The rumen is the largest compartment of ruminant animals. It serves as a "fermenta-
tion room". It is a complicated microbial systems and one of the most fascinating systems
in nature. 400 A wide variety of bacteria, fungi and protozoa act together to bioconvert lig-
nocellulosic plant material into compounds which can be taken up and metabolized by the
ruminant. Ruminal content (fluid and feed material) contains about 1000 microorganisms
per milliliter, including prokaryotic (bacterial and archaeal) and eukaryotic species. 401
399. The word "lampredotto" derives from the Italian word "lampreda", for the resemblance that the
tripe is thought to have to cooked lamprey eel.
400. Weimer P. J., J. B. Russell, R. E. Muck (2009), "Lessons from the cow: what the ruminant ani-
mal can teach us about consolidated bioprocessing of cellulosic biomass", Bioresource Technology, 100:
5323-5331.
401. Qi M, K. D. Jakober, T. A. McAllister (2010), Rumen Microbiology, in R. J. Hudson (Edtr),
Animal and Plant Productivity - Encyclopedia of Life Support Systems (EOLSS). For more information
contact: eolssunesco@gmail.com
106
Animal by-products (ABPs): origins, uses, and European regulations
When ruminant animals are slaughtered the contents of their rumen can become a
viable feed resource and, with appropriate processing, could provide a valuable sources of
useful microorganisms.
Sauer and others 402 suggest intensifying the studies of the ruminal microbial ecosystem
from an industrial microbiologists point of view in order to make use of this rich source of
organisms and enzymes.
Rennet 403 (or Chymosin) is obtained from the stomachs of young mammals, baby cows
or pigs living on milk, and is especially valued when it comes from the inner lining of the
fourth stomach (abomasum) of milk-fed calves. It is a complex of enzymes, 404 used in cheese-
making and one of the first commercially available enzymes used in the food industry. 405
Natural calf rennet is extracted from the inner mucosa of the abomasum of slaughtered
unweaned calves. Of course, these stomach is a by-product of veal production. If rennet is
extracted from older calves (grass or grain fed) the rennet contains less or no chymosin but a
high level of pepsin and can only be used for particular types of milk and cheeses. Milk-spe-
cific rennets are available, such as kid goat rennet for goat's milk and lamb rennet for sheep's
milk, as each ruminant produces a special kind of rennet to digest the milk of its own species.
Ancient Egyptians were the first to discover the use of rennet about 6000 years ago. 406
Since they used the dried stomachs of animals as containers for storing liquids, it is supposed
that the first cheese was occasionally produced when they stored milk in that container. Those
containers could have produced solid chunks (mainly formed by casein) and liquid (whey).
Until 1990, rennet was produced from abomasum, glutinous rice wine, 407 and from
various "vegetable" rennet, some of which were made from the microorganism Mucor mie-
hei. Over recent years, the growth in the cheese industry and the scarcity on calf rennet
have stimulated the research for milk clotting enzyme from alternative sources. 408 Rennet is
produced by genetically engineered bacteria into which the gene for the enzyme has been
inserted.
Caseins comprise the main protein component of milk and are secreted as micelles with
high concentrations of calcium. They are phosphoproteins that represent the products of
four genes equivalent to those that encode the bovine alpha si, alpha s2, beta, and kappa-
402. M. Sauer, H. Marx, D. Mattanovich (2012), "From rumen to industry", Microbial Cell Factories,
vol. 11: 121-124.
403. "Rennet" The Columbia Encyclopedia, 6th ed. 2012. Encyclopedia.com. 21 Feb. 2013. (See:
htmlhttp:// www.encyclopedia.com/).
404. Rennet contains many enzymes, including a proteolytic enzyme (protease) that coagulates the
milk, causing it to separate into solids (curds) and liquid (whey).
405. Fox P. F., P. McSweeney, T. M. Cogan, T. P. Guinee (2004), Cheese: Major cheese groups, Academic
Press., pp. 2.
406. Neelakantan S., A. K. Mohanty, J. K. Kaushik (1999), "Production and use of microbial enzymes
for dairy processing", Current Science, 77 (1): 143-148.
407. Jiang T, L. J. Chen, L. Xue, L. S. Chen (2007), "Study on milk-clotting mechanism of rennet-
like enzyme from glutinous rice wine: proteolytic property and the cleavage site on kappa-casein",
Journal of Dairy Science, 90 (7): 3126-33.
408. Escobar J., S. Barnett (1993), "Effect of agitation speed on the synthesis of Mucor miehei acid
protease", Enzyme and Microbial Technology, 15 (12): 1009-1013; Silveira G. C, G. Monteiro de
Oliveira, E. J. Ribeiro, R. Monti, J. Contiero (2005), "Microbial Rennet Produced by Mucor miehei
in Solid-State and Submerged Fermentation", Brazilian Archives of Biology and Technology, 48 (6):
931-937.
107
Raffaella Leoci
caseins. 409 Fiat and Jolles 410 have studied the structural and physiological aspects of caseins of
various origins. They concluded that if, in the future, "some of the discussed active peptides
(casein fractions of various origins) cannot be characterized in vivo they can all, nevertheless,
be synthesized and used either as food additives or in pharmacology".
4.16 Trachea
Trachea is the scientific name for the windpipe, that is the tube connecting the nose,
mouth, and throat to the lungs. Bovine Cartilage Extract from the trachea is a rich source of:
• Natural chondroitin sulphate (>20%);
• Glycosaminoglycans (also known as mucopolysaccharides);
• Collagen (Type 11 Collagen).
a) Chondroitin sulphate
Chondroitin sulphate (CS) is a compound 411 that is found in blood, cartilage, etc. 412
around joints in the body. Cartilage is a type of connective tissue that is found in the skeletal
systems of many animals.
In 1884, chondroitin sulphate was isolated from cartilage by Krukenberg 413 who de-
scribed the first preparation of glucaminoglycan, but the nature of its monosaccharides and
structure was later described in 1925. In 1940, chondroitin was first identified as a com-
ponent of cartilage. It gained more popularity in 1998 after publication of the book called
"The Arthritis Cure". 414 Since then extensive research has been done to observe the efficacy of
chondroitin sulfate, especially in its combination with glucosamine to treat osteoarthritis 415
or as anti-inflammatory. 416
409. Ginger M. R., M. R. Grigor (1999), "Comparative aspects of milk caseins", Comparative Bio-
chemistry and Physiology - Part B: Biochemistry and Molecular Biology, 124 (2): 133-45.
410. Fiat A. M„ P. Jolles (1989), "Caseins of various origins and biologically active casein peptides and
oligosaccharides: structural and physiological aspects", Molecular and Cellular Biochemistry, 87 (1): 5-30.
411. Chondroitin sulfate is a linear heteropolysaccharide consisting of repeating disaccharide units of
glucuronic acid and galactosamine, which is commonly sulfated at C-4 and/or C-6 of galactosamine.
chondroitin sulfate is a glycosaminoglycan covalently linked to proteins forming proteoglycans. See:
Calabro A., A. Plaas, R. J. Midura, N. J. Goodstone, L. Roden, V. C. Hascall (2000), Structure and
Biosynthesis of Chondroitin Sulphate and Hyaluronan, in R. V. Iozzo (Edtr), Proteoglycans: Structure,
Biology and Molecular Interactions, Marcel Dekker, Inc., New York-Basel, pp.5-26.
412. Blood, Bladder, Cartilage, Fibroblasts, Kidney, Myelin, Nerve Cells, Nervous Tissues, Neuron,
Placenta, Platelet Prostate, Skin, Spleen, Testes, Trachea.
413. Krukenberg C. F. W. (1884), "Die chemischen Bestandtheile des knorpels", Zeitschrift fur Biolo-
gic, 20: 207-326. In a paper published in 1891 by Schmiedeberg, the name of the material was referred
as Chondroitinschwefelsaure, which was derived from chondros (cartilage).
414. Theodosakis J., B. Adderly, B. Fox (1998), The Arthritis Cure: The Medical Miracle That Can
Halt, Reverse, And May Even Cure Osteoarthritis, St. Martin's Griffin Edition, New York.
415. Yves H. (2010), "Advances in the Treatment of Osteoarthritis and the Role of Chondroitin Sul-
phate - A Review", Musculoskeletal Review, 5: 11-17; Kahan A., D. Uebelhart, F. De Vathaire, P. D.
Delmas, J. Y. Reginster (2009), "Long-term effects of chondroitins 4 and 6 sulfate on knee osteo-
arthritis: The study on osteoarthritis progression prevention, a two-year, randomized, double-blind,
placebo-controlled trial", Arthritis & Rheumatism, 60: 524-533.
416. Ronca F., L. Palmieri, P. Panicucci, G. Ronca (1998), "Anti-inflammatory activity of chondroitin
108
Animal by-products (ABPs): origins, uses, and European regulations
Chondroitin sulfate is manufactured from animal sources such as cow cartilage. CS
molecule represents a heterogeneous population the structure of which varies with source.
These polymers are extracted from bovine trachea. Nakano and others 417 have studied an
economical method for its extraction from bovine nasal cartilage "without introducing any
chemicals except acetic acid used for pH adjustment". Chondroitin sulphate seems to have
many applications in pharmaceutical, cosmetic, and food industries.
In pharmaceuticals it has been shown to have chondroprotective 418 and antiatherogen-
ic 419 effects in experimental animals. A chondroitin sulphate-iron complex has been reported
as an antianemic agent, in which CS contributes to an increased bioavailability of iron. 420 In
eye banks, CS is used to increase storage time of corneas 421 and as a solution for preserving
corneas used for transplants and during cataract surgery. In health care medicine applica-
tions, CS is used for the treatment of neuralgia, nerve migraine headaches, joint pain, shoul-
der joint pain, and abdominal pain after surgery. The use of bovine cartilage is promoted
as an alternative treatment for cancer. In veterinary use it can be injected into the joints of
animals to relieve inflammation.
In cosmetics, because of its moisturizing properties and because it possibly increases
elasticity and pliability of skin and has the ability to reinforce skin's intercellular matrix, CS
sulphate", Osteoarthritis Cartilage, 6: 14-21; Mason P. (2007), Dietary Supplements, Third Edition,
Pharmaceutical Press, London-Chicago, pp. 68-70; Egea J., A. G.Garcia, J. Verges, E. Montell, M. G.
Lopez (2010), "Antioxidant, antiinflammatory and neuroprotective actions of chondroitin sulfate and
proteoglycans", Osteoarthritis Cartilage,. 18 (1): S24— S28.
417. Nakano T, N. Ikawa, L. Ozimek (2000), "An economical method to extract chondroitin sul-
phate-peptide from bovine nasal cartilage", Canadian Agricultural Engineering, 42: 205-208.
418. Kalbhen D. A. (1983), "Experimental confirmation of the antiarthritic activity of glycosamino-
glycan polysulfate", Zeitschrift fur Rheumatologie, 42: 178-84; Brennan J. J., E X. Aherne, T. Nakano
(1987), "Effects of glycosaminoglycan polysulfate treatment on soundness, hyaluronic acid content
of synovial fluid and proteoglycan aggregate in articular cartilage of lame boars", Canadian Journal of
Veterinary Research, 51: 394-8; Dean D. D., O. E. Mung, I. Rodriquez, M. R. Carreno, S. Morales,
A. Agudez, M. E. Madan, R. D. Altman, M. Annefeld, D. S. Howell (1991), "Amelioration of lapine
osteoarthritis by treatment with glycosaminoglycan-peptide association complex (Rumalon)", Arthritis
and Rheumatism, 34: 304-313.
419. Morrison L. M., N. L. Enrick (1973), "Coronary Heart Disease: Reduction of Death Rate By
Chondroitin Sulfate a", Angiology, 24 (5): 269-287; Matsushima, T, Y. Nakashima, M. Sugano, H.
Tasaki, A. Kuroiwa, O. Koide (1987), "Suppression of atherogenesis in hypercholesterolemic rabbits
by chondroitin-6-sulfate", Artery, 14 (6): 316-337; Kevin Jon W. (2001), "Arterial wall chondroitin
sulfate proteoglycans: diverse molecules with distinct roles in lipoprotein retention and atherogenesis",
Current Opinion in Lipidology, 12 (5) : 477-487.
420. Fochi E, M. Ciampini, G. Ceccarelli (1985), "Efficacy of iron therapy: a comparative evaluation
of four iron preparations administered to anaemic pregnant women", The Journal of International Med-
ical Research, 13 (1): 1-11; Barone D., L. Orlando, E. Vigna, S. Baroni, A. M. Borghi (1988), "Ferric
chondroitin 6-sulfate (Condrofer): a new potent antianaemic agent with a favourable pharmacokinetic
profile", Drugs under Experimental and Clinical Research, 14 (1): 1-14.
421. Keates R. H., B. Rabin (1988), "Extending corneal storage with 2.5% chondroitin sulfate (K-
Sol)", Ophthalmic Surgery, 19 (11): 817-820; Lass J. H., W. J. Reinhart, D. L. Skelnik, W. E. Bruner,
R. P. Shockley, J. Y. Park, D. L. Horn, R. L. Lindstrom (1990), "An in vitro and clinical comparison of
corneal storage with chondroitin sulfate corneal storage medium with and without dextran", Ophthal-
mology, 97 (1): 96-103; Tachibana A., M. Sawa (2002), "Development of novel corneal storage me-
dium: first report. Examinations of rabbit cornea", Japanese Journal of Ophthalmology, 46 (4): 377-83.
109
Raffaella Leoci
is used in eye lotions, in preparations for the preservation of eyesight, in eye drops for dry
eyes, and in shampoos and skin creams. 422
In the food industries it is used to prepare mayonnaise and dressings, 423 as a nutraceuti-
cal 424 in health food drinks (CS is approved by the FDA for these uses). 425
Chondroitin sulfate is often sold in combination products that also contain glucosa-
mine sulfate. So far, there is no evidence that the combination products work any better than
either chondroitin sulfate or glucosamine sulfate alone.
Some doubt on the effectiveness of CS in all the above-mentioned applications was
expressed by Henrotin and others 426 : "Despite the moderate effects of CS on pain and func-
tion, CS is an interesting product for the management of knee osteoarthritis. Clinical evi-
dence is in favor of a slow-acting effect on symptoms in moderate knee osteoarthritis. CS
is recommended by the most popular guidelines. Its safety profile is surely one of its main
benefits for the treatment of aging patient with some comorbidity".
However, there isn't enough information to know if chondroitin sulfate is effective for
other conditions for which people use it: heart disease, osteoporosis (weak bones), and high
cholesterol. Wandel and others 427 in their research to determine the effect of glucosamine,
chondroitin, or the two in combination on joint pain and on radiological progression of
disease in osteoarthritis of the hip or knee conclude that "Compared with placebo, glucosa-
mine, chondroitin, and their combination do not reduce joint pain or have an impact on
narrowing of joint space. Health authorities and health insurers should not cover the costs
of these preparations, and new prescriptions to patients who have not received treatment
should be discouraged". Clegg and others 428 confirm the ineffectiveness of glucosamine and
chondroitin sulfate alone or in combination for the treatment of osteoporosis because they
do not reduce pain effectively in patients with osteoarthritis of the knee. Exploratory analy-
ses suggest that the combination of glucosamine and chondroitin sulfate may be effective in
the patients with moderate to severe knee pain.
422. Nakamura M., M. Hikida, T. Nakano (1992), "Concentration and molecular weight dependency
of rabbit corneal epithelial wound healing on hyaluronan", Current eye Research, 11 (10): 981-986.
423. Yabe Y., T. Ninomiya, H. Kashiwaba, T. Tatsuno, T. Okada (1987), "Determination of sodium
chondroitin sulfate added in foods", Journal of the Food Hygienic Society of japan, 28 (1): 13-18; Ha-
mano T., Y. Mitsuhashi, N. Acki, S. Yamamoto (1989), "High-performance liquid chromatographic
assay of chondroitin sulphate in food products", Analyst, 114 (8): 891-893.
424. Kalra E. K. (2003), "Nutraceutical - definition and introduction", AAPS PharmSci (American
Association of Pharmaceutical Scientists), 5 (3): 27-28.
425. Tallon M. J. (2007), Key Trends in Nutraceutical Food and Drinks, Business Insights Ltd., Not-
tinghamshire (UK), p. 110.
426. Henrotin Y, M. Mathy, C. Sanchez, C. Lambert (2010), "Chondroitin Sulfate in the Treatment
of Osteoarthritis: From in Vitro Studies to Clinical Recommendations", Therapeutic Advances in Mus-
culoskeletal Disease, 2 (6): 335-348.
427. Wandel S., P. Jiini, B. Tendal, E. Nuesch, P. M. Villiger, N. J. Welton, S. Reichenbach, S. Trelle
(2010), "Effects of glucosamine, chondroitin, or placebo in patients with osteoarthritis of hip or knee:
network meta-analysis", British Medical Journal, 341: c4675
428. Clegg D. O. and others (2006), "Glucosamine, chondroitin sulfate, and the two in combination
for painful knee osteoarthritis", The New England Journal of Medicine, 354 (8): 795-808.
110
Animal by-products (ABPs): origins, uses, and European regulations
According to Lauder 429 "while the safety of CS is not presently in doubt, poor quality
finished products have the potential to compromise clinical and lab-based studies and will
fail to give consumers all of the benefits available". In conclusion Volpi's 430 position on "a
need for specific and accurate analytical procedures, which should be enforced to confirm
purity and label claims both for raw materials and finished chondroitin sulfate products, and
also to govern the origin of ingredients" seems to be correct.
429. Lauder R. M. (2009), "Chondroitin sulphate: A complex molecule with potential impacts on a
wide range of biological systems", Complementary Therapies in Medicine, 17 (1): 56—62.
430. Volpi N. (2009), "Quality of different chondroitin sulfate preparations in relation to their thera-
peutic activity", Journal of Pharmacy and Pharmacology, 61 (10): 1271-80.
Ill
Raffaella Leoci
112
Animal by-products (ABPs): origins, uses, and European regulations
PART II
Treatment and legislation on ABPs and waste
113
Raffaella Leoci
114
Animal by-products (ABPs): origins, uses, and European regulations
5. Collection and treatment plants for ABPs
It has already been mentioned that wastes typically arise from food processing and
manufacturing plants, distribution premises, food markets, wholesale and retail outlets, and
catering facilities (including household kitchens). The European Community has issued a
number of directives for waste management.
Landfilling of bio-waste is addressed in the Landfill Directive 431 that requires the diver-
sion of biodegradable municipal waste from landfills. The Industrial Emissions Directive 432
lays down the main principles for the permitting and control of waste treatment installa-
tions. The incineration of bio-waste is regulated in the Waste Incineration Directive, 433 while
the health rules for composting and biogas plants that treat animal by-products are laid
down in the ABPs Regulation. 434
Since the EU banned the use of rendered animal proteins from the feed chain in 1999,
the costs for the treatment of ABPs have increased considerably. Since that EU ABP-Reg-
ulation, new opportunities for the utilization of slaughterhouse wastes were opened. For
example, in the agri-industrial sector some EU biogas plants 435 are now using pasteurized
ABP as co-substrates together with manure, rumen content, and other energy crops (blood,
minced hind gut, fat, etc) .
There are many premises that generate, store, use, handle and dispose of animal by-
products, as set out below:
5.1 - Rendering plants: handling and storage plants;
5.2 - Incinerators/Co-incineration Plants;
5.3 - Landfill;
5.4 - Anaerobic digestion (biogas) plants;
5.5 - Composting;
5.6 - Petfood plants.
5.1 Rendering plant: handling and storage plants (previously known as intermediate plants)
Not all ABP are sent to rendering plants. As already said, meat rich by-products like
lungs, liver and hearts are often sold to wet pet food plants and to other utilizations. Ren-
431. Council Directive 1999/31 /EC of 26/4/99 on the landfill of waste (Official Journal of the Euro-
pean Communities L 182 of 16.7.99).
432. Directive 2010/75/EU of the European Parliament and of the Council of 24 November 2010 on
industrial emissions (integrated pollution prevention and control) (Official Journal of the European
Union L 334 of 17.12.2010).
433. Directive 2000/76/EC of the European Parliament and of the Council of 4 December 2000 on
the incineration of waste (Official Journal of the European Union L 332 of 28.12.2000).
434. Regulation (EC) No 1069/2009 of the European Parliament and of the Council of 21 October
2009 laying down health rules as regards animal by-products and derived products not intended for
human consumption and repealing Regulation (EC) No 1774/2002 (Animal by-products Regulation)
(Official Journal of the European Union L 300 of 14.1 1.2009).
435. Kirchmayr R., R. Scherzer, D. L. Baggesen, R. Braun, A. Wellinger (2003), Animal by-Products
and Anaerobic Digestion - Requirements of the European Regulation (EC) No 1774/2002, IEA Bio-
energy, Task 37 - Energy from Biogas and Landfill Gas - In Cooperation with BIOEXELL - Biogas
Center of Excellence.
115
Raffaella Leoci
dering is a process which involves cooking animal carcasses or ABPs at high temperature,
sometimes under pressure, allowing water to be removed and tallow to be separated from the
proteinaceous material. Rendering reduces the volume of disposable material.
The process involves the crushing and grinding of ABP, followed by heat treatment to
reduce the moisture content and kill micro-organisms. Heating separates the tallow from
the bone and protein. The first can be used as a fuel for the process. The protein and bone
are made into meat and bone meal. In research Ramirez and others 436 determined that the
weighted average proportion of thermal process energy derived from tallow for six plants in
UK, between 2006 and 2008, was 76% and the remaining 24% being derived from natural
gas.
Separation of materials is vital to ensure that the risks associated with ABPs are ef-
fectively controlled. This includes keeping raw feedstock separate from the processed final
product and also allows separation of by-product categories . Procedures must be followed
to ensure categories of animal by-products are kept separate from each other at all times in
order to minimize the risk of cross-contamination.
In addition, to prevent by-pass of material within facilities, p rocedures should be
adopted to control the movement of personnel between areas. . Color coded protective
clothing ensures separation between personnel working in different areas (clean and un-
clean). Procedures must also be adopted to control by-pass risks in communally used areas
such as lavatories, changing rooms, and canteens. All staff working on the premises must be
adequately trained for their job.
The method of required processing will depend on the Category and nature of the raw ma-
terial being processed. All rendering plants would contain at least the following process steps:
1. Reception: where the incoming ABPs enter the facility, usuallya form of double
door , that provides an odor lock to prevent the release of odors during unloading
operations;
2. Identification of biological hazards: the biological hazards (human and animal
health) should be identified in view of origin of the raw material;
3. Storage and handling: ABPs are stored, usually in lidded hoppers, for the shortest
possible time. Usually ABP boxes are identified by distinctive colors, depending on
ABP Category, in accordance with Regulation 1069/2009 of the European Parlia-
ment. To comply with ABPs regulations, the raw material must be accompanied by
official records of its kind, when received, and when processed;
4. Size reduction: before being passed into the rendering process the ABPs need to be
pulpedin order to ensure that all parts of the rendered materials receive sufficient
treatment. By mashing a uniform particle size is achieved that equally distributes
raw mass. Processing: the crushed ABPs are heated in the rendering plant, where a
defined temperature and residence time allow sterilization and the degradation of
cellular links for the release of fat;
436. Ramirez A. D., A. Humphries, S. L. Woodgate, R. W. Wilkinson (2012), The potential for en-
ergy self-sufficiency in the United Kingdom rendering industry, in Brebbia C. A., S. S. Zubir (Edtrs),
Management of Natural Resources, Sustainable Development and Ecological Hazards, WIT Press, South-
ampton (UK), pp. 483-94.
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Animal by-products (ABPs): origins, uses, and European regulations
5. Separation of the solid/liquid fraction: the rendering mass is put into the decant-
ers where the liquid fraction is separated from the solid fraction. The solid fraction
contains the protein mass of tissue, while the liquid fraction contains fat and water;
6. Separation of the fat: after leaving the decanter, the liquid fraction is lead to an interme-
diate storage container where the components are separated by a centrifugal separator.
That centrifugal force separates the liquid fraction into fats, wastewater, and sludge;
7. Storage of processed materials: meal and fat manufactured by rendering are stored
pending users. After treatment the final fat can be used for biodiesel production or
can fuel a combustion unit;
8. Washing and cleaning: the plant building, processing equipment, transportation
vehicles, and ABP delivery equipment require regular washing and cleaning;
9. Treatment of odorous emissions: because rendering produces odors, plants must
have extraction equipment for effective odor abatement;
10. Effluent treatment: all effluent produced must be treated in an approved process on
site or sent off site for treatment according to the laws.
As we will see later, Annex IV chapter III of the EU Commission Regulation provides
detail of the seven standard processing methods including time, temperature and particle
size requirements.
5.2 Incinerators/Co-incineration Plants
In the European Union, incinerators and co-incinerators burning ABPs must be operat-
ed in accordance with "Waste Incineration Directive (Directive 2000/76/EC)", "Regulation
(EC) No 1069/2009", and "Commission Regulation (EU) No 142/2011 of 25 February
201 1" 437 (as amended by Commission Regulation (EU) No 749/2011 of 29 July 201 1 438 ),
which require the disposal or recovery of the resulting ash to be carried out in accordance
with environmental legislation. To minimize risk to animal health, Regulation 142/2011
requires that animals are prevented from having access to the ash.
Ash from the incineration or co-incineration of ABPs is subject to waste controls and is
normally disposed of in a permitted landfill. Where the feedstock to the incinerator and co-
incineration plant is restricted to either animal carcasses or poultry litter only, the completely
combusted ash can be spread on land to provide nutrients.
Annex III Section 2 of the EU Commission Regulation states: Operating conditions
-Incineration or co-incineration plants will be designed, equipped, built and operated in
such a way that the exhaust gas, resulting from the process, is held even under the most
unfavourable conditions, at a minimum of 850°C for 2 seconds or 1 100°C for 0.2 seconds,
as measured at a representative point of the chamber, where the incineration or the co-
incineration is carried out. The temperature of a minimum of 850°C for 2 seconds is often
achieved using a secondary chamber. The gas temperature in the secondary chamber must
be achieved before by-product incineration begins.
437. Commission Regulation (EU) No 142/2011 of 25 February 2011, "implementing Regulation
(EC) No 1069/2009 of the European Parliament and of the Council laying down health rules as re-
gards animal by-products and derived products not intended for human consumption and implement-
ing Council Directive 97/78/EC as regards certain samples and items exempt from veterinary checks
at the border under that Directive", Official Journal of the European Union, L 54 on 26.02.201 1.
438. Official Journal No L 198 on 30.7.2011.
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Raffaella Leoci
Incineration and combustion plants for organic material, such as waste with energy-
recovery, are common. All new plants in OECD countries must meet strict emission stand-
ards, including those on nitrogen oxides (NOx), sulphur dioxide (S02), heavy metals, and
dioxins. Incineration of ABPs must take place at an approved plant. With or without energy-
generation, incineration of ABPs must be approved under the "Directive on the Incineration
of Waste 2000/76 /EC" . Among the many provisions the following are important to recall.
(We will revisit these subjects in the next section).
In order to prevent the contamination of foodstuffs with pathogenic agents, establish-
ments or plants processing animal by-products should operate on a separate site from slaugh-
terhouses or other establishments in which foodstuffs are processed. (Commission Regula-
tion (EU) No 142/2011, p. 14, page 5).
For the promotion of science and research and to ensure the best possible use of ani-
mal by-products and of derived products in the diagnosis of human or animal diseases,
the competent authority should be authorized to lay down conditions for samples of such
materials for research, educational and diagnostic purposes. (Commission Regulation (EU)
No 142/2011, p. 19, page 6)
Pursuant to Regulation (EC) No 1069/2009, operators are to ensure that animal by-
products and derived products are traceable at all stages of the chain of manufacturing, use
and disposal, so as to avoid unnecessary disruptions of the internal market in the case of
events which are linked to actual or potential risks to public or animal health. Traceabil-
ity should therefore not only be ensured by operators generating, collecting or transport-
ing animal by-products, but also by operators disposing of animal by-products or derived
products, by incineration, co-incineration or landfilling. (Commission Regulation (EU) No
142/2011, p. 22, page 7)
Certain imported materials for the production of pet food should be handled and used
under conditions that are appropriate to the risk that such materials may pose. In particular,
provision should be made for their safe channeling to establishments or plants of destination
where such materials, as well as Category 3 material, are incorporated into pet food. (Com-
mission Regulation (EU) No 142/2011, p. 28, page 8)
Apiculture by-products which are to be placed on the market should be free of certain
diseases to which bees are susceptible. These diseases are listed in Council Directive 92/65/
EEC of 13 July 1992 which establishes animal health requirements governing trade in and
imports into the community of animals. Semen, ova and embryos not subject to animal
health requirements laid down in specific Community rules are referred to in Annex A (I)
to Directive 90/425/EEC (Commission Regulation (EU) No 142/2011, p. 34, page 11).
If a premises houses which has livestock and only by-products arising on that premise
are incinerated then there must be physical and operational separation such that the livestock
cannot gain access to the incinerator and there is no risk of passive transfer of by-products to
livestock by personnel or equipment. This will require dedicated equipment and clothing for
use when operating the incinerator or cleansing and disinfection prior to use associated with
livestock (Commission Regulation (EU) No 142/2011, Annex III, Chapter I, Section I).
If an incinerator will be used to incinerate by-products from more than one premise,
it must be located on a site that has no livestock: a separate premise. For the location to be
considered a separate premise it must, as a minimum, have a defined secure boundary and
dedicated entrance . It cannot be incorporated into , or be a part of, i a livestock premise.
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Animal by-products (ABPs): origins, uses, and European regulations
It may be possible to approve the use of a mobile incinerator 439 at specified locations
which otherwise meet the regulatory requirements of approval for a static incinerator. In
such cases an agreed procedure for the cleansing and disinfection of all mobile equipment
prior to movement to a new location will be a condition of any approval issued.
Article 24 of the "Approval of establishments or plants" (Regulation (EC) No 1069/2009)
establishes that operators will ensure that establishments or plants under their control are
approved by competent authority, where such establishments or plants carry out disposal,
as waste, by incineration or co-incineration of animal by-products and derived products,
excluding establishments or plants which have a permit to operate in accordance with Direc-
tive 2000/76/EC.
This applies to most incinerators on farms, at hunt kennels and knackers' yards, and at
pet crematoria. Those incinerator plants that burn "processed products" (e.g. meat and bone
meal, tallow), catering waste or material other than animal carcasses/parts of carcasses must
meet Directive 2000/76/EC requirements.
Regulation (EC) No 142/2011 (Chapter III) permits low capacity (< 50kg/hour) in-
cinerator plants, that are equipped with an auxiliary burner, to burn the following Speci-
fied Risk Material: Category 1 materials referred to in Article 8 (b), (e) and (f), Category
2 materials referred to in Article 9 or Category 3 materials referred to in Article 1 0 of the
Regulation (EC) No 1069/2009, that we will see ahead.
5.3 Landfill
Nowadays the production of methane from biodegradable waste decomposing in land-
fills constitutes a threat to the environment. The EU Landfill Directive (1999/31 /EC) has
obliged Member States to gradually reduce the amount of biodegradable waste that they add
to landfills from 1995 levels until 2016 (for some countries till 2020).
The Landfill Directive does not prescribe specific treatment options for the diverted
waste. The benefits of proper bio-waste management, besides avoided emissions of methane
and carbon dioxide, would be the production of compost that contributes to enhanced soil
quality and methane that is a valuable source of energy. However, Member States are often
inclined not to opt for composting or bio-gas production but for the easiest and cheapest
option, which is incineration or landfilling.
5.4 Anaerobic digestion
In recent decades soaring globalization, large-scale livestock production, and an escala-
tion in epizootic diseases, has increased the need for biosecurityso that the transmission of
disease to the food chain can be minimize. The potential negative environmental impact of
biological waste, and waste from ABPs, can be minimized by recycling them into plant nu-
trients (compost or fertilizers) or converting them into biogas. Since biological waste and/or
ABPs can inadvertently spread infectious diseases, obtaining general acceptance for hygieni-
cally safe end-products is needed.
There is no doubt that landfilling is the worst waste management option for bio-waste
and ABPs. To encourage the management of biodegradable waste diversion from landfills,
there seems to be several environmentally favorable options. While a waste management
439. Commission Regulation (EU) No 142/2011, Annex I, pag. 36.
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Raffaella Leoci
hierarchy applies to the management of bio-waste, in specific cases there may be justifica-
tion for departures since the environmental balance of the various options available for the
management of this waste depends on a number of local factors, i. e. on collection sys-
tems, waste composition and quality, climatic conditions, and the potential of use of various
waste-derived products such as electricity, heat, methane-rich gas or compost. Therefore,
national strategies for the management of this waste should be based on a structured and
comprehensive approach (e.g. Life Cycle Assessment, Life Cycle Thinking, ecc). Anaerobic
digestion and composting treatment seem the suitable treatment methods that can combine
biosecurity aspects with environmental, economic and nutrient recycling aspects, in order
to create a beneficial whole-farm approach. 440 Composting and biogas plants may also be
approved by an alternative method set out in Regulation 208/2006. Under this Regulation,
approvals are issued according to satisfactory demonstration of sufficient pathogen kill in
the treatment process.
As already pointed, in the European Union the landfilling of bio-waste is regulated in
the Landfill Directive (1999/31/EC), while the rules for composting and biogas plants that
treat animal by-products are regulated in the ABPs Regulation.
It is necessary to remember some definitions of terms.
"Organic fertilizers" can be defined as materials of animal origin used to maintain or
improve the physical makeup, chemical properties, and biological activities of soils and/or
plant nutrition. Organic fertilizers contain derived products or processed animal proteins
(PAP) such as bloodmeal, feathermeal, fishmeal, former foodstuffs and meat and bone meal;
"Soil improvers" or "compost" can be defined as materials of vegetal and/or animal ori-
gin used to maintain or improve the physical and biological activities of soils. They cannot
be defined "fertilizers";
"Bio-waste" can be defined as food and kitchen waste from households, restaurants,
caterers and retail premises; comparable waste from food processing plants; waste from ABPs
or processing ABPs; and biodegradable garden and park waste. It does not include agricul-
tural residues or forestry, manure, sewage sludge and by-products of food production that
never become waste;
According to the European Union, "Fertilizer" means material of which the main func-
tion is to provide nutrients for plants 441 . That definition was absorbed by the Italian law. 442
The "Concimi" (Fertilizers) are divided into «fertilizers CE» e «national fertilizers", whose
types and characteristics are reported respectively in Annex I of the Regulation (CE) n.
2003/2003 and Article 2 of Italian Legislative Decree 2010, 29 April, n.75;
"Soil improvers" (ammendanti): materials to be added to the soil in situ, primarily to
maintain or improve its physical and/or chemical and/or biological activity (Article 2, Italian
Legislative Decree 2010, 29 April, n.75).
440. Albihna A., B. Vinneras (2007), "Biosecurity and arable use of manure and biowaste — Treat-
ment alternatives", Livestock Science, 1 12 (3): 232-239.
441. Article 2 of the Regulation (EC) No 2003/2003 of the European Parliament and of the Coun-
cil of 13 October 2003, relating to fertilizers (Official Journal of the European Union, L 304 on
21.11.2003).
442. Article 2 of the Italian Legislative Decree n.75, 2010, 29 April.
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Animal by-products (ABPs): origins, uses, and European regulations
Biogas plants
Biogas is a biofuel. Generally biogas refers to the gas produced from organic matter as
it is broken down by biological means, in the absence of oxygen. 443 Therefore, biogas can
be produced, using anaerobic digesters and by the digestion or fermentation of biodegrad-
able materials such as biomass, food waste, manure, sewage sludge, ABPs, municipal waste,
green waste and energy crops such as cereals silage. Produced biogas is comprised primar-
ily of methane (about 58-60%) and carbon dioxide (about 38-42%) and may have small
amounts of moisture, nitrogen, hydrogen sulphide, and siloxanes. 444 Natural gas, as known,
is composed of about 97% methane. Technologies such as pressure swing absorption and
water scrubbing are used to remove C0 2 from the biogas stream, converting it to renewable
natural gas. The methane can be used for heating, production of electricity, and many other
machines that use an internal combustion engine.
Biogas plants are systems that use a bacteriological process called anaerobic digestion
to convert organic waste into biogas. 445 According to Commission Regulation (EU) No
142/201 1, ANNEX I, "biogas plant means a plant in which animal by-products or derived
products are at least part of the material which is submitted to biological degradation under
anaerobic conditions".
Usually, the biogas plant consists of two components: a fermentation tank or digester
and a gasholder. The first is a cube-shaped or cylindrical waterproof container with an inlet
into which the fermentable mixture is introduced as a liquid slurry or in solid materials
form. Bacteria within the digester tank breaks down the waste and, as it decomposes, gases.
Through a pressurized system, the gasholder conducts the flow of these gases upward into
a hole that is normally an airproof container that cuts off air to the digester and collects
the generated gas. 446 A solid residue (digestate), similar but not identical to compost, and a
liquid liquor remain. The digestate can be separated out, composted, and sold . 447 The liquor
can be used as a fertilizer.
Several anaerobic digester technologies exist. Each is designed to process specific waste
streams. Nowadays, there are two main categories of anaerobic digesters: liquid digesters and
solid digesters. The first are systems in which the substrate inside the digester is adequately
fluid to be pumped (less than 15% dry matter). Solid digesters are systems where the mate-
rial inside the digester remains solid and is expelled in a solid form. They may run in batches
or continuously or semi-continuously. 448
443. Torien D. E, W. H. J. Hattingh, J. P. Kotze, P. G. Uriel, W. A. Pretorius, G. G. Cillie, M. R,
Henzen, G. J. Stander, R. D. Baillie (1969), "Anaerobic Digestion - Review Paper", Water Research,
Pergamon Press, 3: 385-416.
444. Wolfe R.S. (1971), "Microbial Formation of Methane", Advances in Microbial Physiology, 6:107-146.
445. Gunaseelan V. N. (1997), "Anaerobic digestion of biomass for methane production: a review",
Biomass and Bioenergy, 13:83-114.
446. Biswas T. D. (1977), "Biogas Plants: Prospects and Limitations", Invention Intelligence, 12: 71-78.
447. The value of digestates as fertilizers depends on the make-up of the digestate, and its classifica-
tion in waste terms. When harmful inorganic substances (e.g. heavy metals), organic substances (e.g.
medicinals, PAHs, PCBs, PCDDs, etc.) or injurious weed seeds are present, land application of the
digestate may not be possible.
448. Alvarez R., G. Liden (2008), "Semi-continuous co-digestion of solid slaughterhouse waste, ma-
nure, and fruit and vegetable waste", Renewable Energy, 33 (4), 726-734.
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Raffaella Leoci
For decades anaerobic digestion has been used for wastewater treatment and for stabi-
lization and volume reduction of sewage sludge, animal manure and organic wastes diges-
tion. Experience has shown that the spreading of digested material of this wastewater type
has not caused health problems.When correct processing methods are followed, anaerobic
digestion provides a successful method for accessing the energy and nutrient content con-
tained in organic material. Consequently, the utilization of anaerobic digestion for organic
waste management permits a significant movement up the waste hierarchy over other
management methods.
The value of digestates as fertilizers depends on the make-up of the digestate and its
classification in waste terms. With correct storage and application methods and sound agri-
cultural practices, the risk of volatilization and runoff of nutrients of digested material can
be greatly reduced compared to risks of storage and application of untreated organic waste
and manure.
Recently anaerobic digestion has received increasing attention as a mainstream energy
conversion process because it is based on renewable agricultural biomass (energy crops), as
well as on co-digestion of various industrial by-products, including animal by-products and
wastes. 449 By converting waste into energy, biogas plants reduce odors and pathogens, pro-
duce an enhanced fertilizer and reduce greenhouse gas emissions.
Worldwide, Germany is the market leader in this field, with over 4,000 on-farm anaero-
bic digesters generating more than 1,200 MW of clean power (German Biogas Association,
www.biogas.org).
Sami Luste and other 450 have studied the effect of five pre-treatments (thermal, ultra-
sound, acid, base and bacterial product) on hydrolysis and methane production potentials
of four by-products from meat-processing industry. In batch experiments, thermal treatment
increased methane production potential of drumsieve waste, acid of grease trap sludge and
all pre-treatments of dissolved air flotation sludge. However, with all other pre-treatments,
methane production potential was decreased compared to untreated materials, apparently due
to inhibition by hydrolysis products and/or possible re-crystallization of some compounds.
5.5 Composting
Natural composting, or biological decomposition, began with the first dead plants on
earth and has been going on ever since. 451 As vegetation falls to the ground, it slowly decays,
providing minerals and needed nutrients for plants, animals, and microorganisms.
Compost is organic material that can be used as a soil amendment or as a medium to
grow plants. Mature compost is a stable material with a content called humus that is dark
brown or black and has a soil-like, earthy smell. It is created by combining organic wastes
(e.g., yard trimmings, food wastes, manures, etc.) in proper ratios into piles, rows, or vessels.
449. Braun R., R. Kirchmayr (2003), Implementation Stages of Directive EC 177412002 on Animal
Byproducts, inT. Al Seadi and J. Bo Holm-Nielsen (Edtrs), The Future of Biogas in Europe II -European
Biogas Workshop, October 2-4, 2003, University of Southern Denmark Esbjerg/Denmark.
450. Luste S., S. Luostarinen, M. Sillanpaa (2009), "Effect of pre-treatments on hydrolysis and meth-
ane production potentials of by-products from meat-processing industry" ', Journal of Hazardous Ma-
terials, 164 (1): 247-255.
451. Garland G. A., T. A. Grist, R. E. Green (1995), "The compost story: from soil enrichment to
pollution remediation", Biocycle, 36 (10): 53-56.
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Animal by-products (ABPs): origins, uses, and European regulations
Composting benefits are the following:
• Soil conditioner. Using compost, rich humus for lawns and gardens are created and the
compost enrichment helps retain moisture in the soil.
• Recycles kitchen and yard waste. Composting can convert as much as 30% of house-
hold waste into soil conditioner and growing mediums.
• Introduces beneficial organisms to the soil. Microscopic organisms in compost help
aerate the soil, break down organic material for plant use and ward off plant disease.
• Good for the environment.
• Reduces landfill waste. Most landfills in Europe are quickly filling up; many have al-
ready closed down.
In a composting plant animal by-products, or derived products , makeup at least part of
the material submitted to biological degradation under aerobic conditions. 452
It is best not to compost fish scraps (they will attract pests), perennial weeds (they can
be spread with the compost), or diseased plants. Pet manures are not included in compost
that will be used on food crops. Peach peels, banana peels, and orange rinds may contain
pesticide residue; black walnut leaves should not be composted since they can contain tox-
ins. Sawdust may be added to the compost, if it is clean and contains with no lubricating oil
residue from cutting equipment.
A new compost technology, known as compost bioremediation, is currently being used
to restore contaminated soils, manage stormwater, control odors, and degrade volatile or-
ganic compounds. 453
5.6 Pet food plants
Dogs and cats are carnivores, and therefore consume meat-based diet. The protein used
in pet food comes from a variety of sources. When cattle, swine, chickens, lambs, etc., are
slaughtered, lean muscle tissue is trimmed away from the carcass for human consumption.
About 50% of the weight of every slaughter house animal does not directed to human food.
Whatever remains of the carcass (ABPs) can be used in pet food, animal feed, fertilizer, and
for others uses. Using Category 3 materials 454 of Regulation (EC) No 1069/2009, which are
452. Annex I - Definitions as referred to in article 2 of Commission Regulation (EU) No 142/2011.
453. To know more about see: United States Environmental Protection Agency, Innovative Uses of
Compost Bioremediation and Pollution Prevention, Solid Waste and Emergency Response (5306W),
EPA530-F-97-042 October 1997 (www.epa.gov); Buyuksonmez E, R. Rynk, T. E Hess, E. Bechinski
(1999), "Occurrence, Degradation, and Fate of Pesticides During Composting Part I: Composting,
Pesticides, and Pesticide Degradation", Compost Science and Utilization, 7 (4), 66-82; Buyuksonmez
F., R. Rynk.T. F. Hess, E. Bechinski (2000), "Occurrence, Degradation, and Fate of Pesticides During
Composting, Part II: Occurrence and Fate of Pesticides in Compost and Composting Systems", Com-
post Science and Utilization, 8(1), 61-81; Buyuksonmez F., S. Sekeroglu (2005), "Presence of pharma-
ceuticals and personal care products (PPCPs) in biosolids and their degradation during composting",
Journal of Residuals Science & Technology, 2 (1), 31-40.
454. As we will see ahead, briefly they are: a) carcases and parts of animals slaughtered or, in the case of
game, bodies or parts of animals killed, and which are not intended for human consumption for com-
mercial reasons; b) carcases and the parts originating either from animals that have been slaughtered in
a slaughterhouse and were considered fit for human consumption (heads of poultry, hides and skins,
including trimmings and splitting thereof, horns and feet, including the phalanges and the carpus
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materials from healthy animals that would be fit for human consumption but are not con-
sumed for reasons of culture and customer choice are forwarded to specialized industries that
produce chilled, frozen and dried ingredients for the meat and poultry sectors.
According to Annex I of the Commission Regulation (EU) No 142/2011, 'Pet food'
means feed for pet animals and dog chews that:
• contain Category 3,
• may contain imported Category 1 material455 comprising of animal by-products
derived from animals which have been submitted to illegal treatment as defined in
Article l(2)(d) of Directive 96/22/EC or Article 2(b) of Directive 96/23/EC.
More simply, pet foods are plants or animal materials intended for consumption by pets.
Rendering is a process that converts waste animal tissue into value-added materials. Render-
ing can be carried out on an industrial, farm, or kitchen scale. Of course, pet food industry
is an extension of the human food and agriculture industries. Pet food provides a convenient
way for slaughterhouse offal, grains considered "unfit for human consumption," and similar
waste products to be turned into profit. Catering waste (including catering waste processed
in an ABP rendering plant) cannot be used in the manufacture of pet food.
It is known that also the following foods are potentially unsafe for cats and dogs 456 :
• Allium 457 : the toxic principles present in them causes the transformation of hemoglobin
into methemoglobin, consequently resulting in hemolytic anemia;
• Raisins and grapes: large doses of raisins can be poisonous to pets and can cause
vomiting, diarrhea, lethargy, abdominal pain, lack of appetite and kidney damage;
• Macadamia nuts and walnuts: can cause vomiting, lethargy, hyperthermia, abdominal
and metacarpus bones, tarsus and metatarsus bones, pig bristles, feathers); c) animal by-products from
poultry and lagomorphs slaughtered on the farm, which did not show any signs of disease communi-
cable to humans or animals; d) blood; e) animal by-products arising from the production of products
intended for human consumption; f) products of animal origin, or foodstuffs containing products
of animal origin, which are no longer intended for human consumption for commercial reasons; g)
petfood and feedingstuffs of animal origin, or feedingstuffs containing animal by-products or derived
products, which are no longer intended for feeding for commercial reasons; h) blood, placenta, wool,
feathers, hair, horns, hoof cuts and raw milk; i) aquatic animals, except sea mammals, which did not
show any signs of disease; j) material originating from animals which did not show any signs of disease
(shells, hatchery by-products, eggs, egg by-products, including egg shells); k) aquatic and terrestrial in-
vertebrates; i) hides and skins, hooves, feathers, wool, horns, hair and fur originating from dead animals
that did not show any signs of disease; 1) hides and skins, hooves, feathers, wool, horns, hair and fur
originating from dead animals that did not show any signs of disease; m) adipose tissue from animals.
455. They are briefly: a) entire bodies and all body parts, including hides and skins of animals sus-
pected of being infected by aTSE, animals other than farmed and wild animals, including in particular
pet animals, zoo animals and circus animals, animals used for experiments, animal by-products derived
from animals which have been submitted to illegal treatment, containing residues of other substances
and environmental contaminants.
456. Gault G., P. Berny, G. Lorgue (1995), "Plants which are toxic for pets", Recueilde medecine veteri-
naire, vol. 171 (2-3): 171-176; Kovalkovicova N., I. Sutiakova, J. Pistl, V. Sutiak (2009), "Some food
toxic for pets", Interdisciplinary Toxicology, 2 (3): 169-76.
457. Salgado B. S., L. N. Monteiro, N. S. Rocha (201 1), "Allium species poisoning in dogs and cats",
Journal of Venomous Animals and Toxins including Tropical Diseases, vol. 17 (1): 4-11; Cope R. B.
(2005), "Allium species poisoning in dogs and cats", Veterinary Medicine, 100 (8): 562-6.
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Animal by-products (ABPs): origins, uses, and European regulations
pain, stiff joints, lameness and tremors;
• Garlic (in large doses) : can cause gastrointestinal problems such as vomiting and diarrhea.
• Onions 458 : damages hemoglobin and is cumulative. They contain a substance
(N-propyl disulphide) that destroys red blood cells in the cat;
• Chocolate, coffee-based products 459 : can cause irregular heart rate and rhythm, restless-
ness, hyperactivity, diarrhea, vomiting, panting, muscle tremors, abdominal pain,
bloody urine, increased body temperature, seizures, coma and possibly even death.
Theobromine, a component of chocolate, is a toxic compound. Caffeine 46 " is also pre-
sent in chocolate and a toxic component, but in much smaller amounts than Theobro-
mine. Both Theobromine and Caffeine are members of a drug class called Methylxanines;
• Alcoholic beverages: any type of alcohol can be poisonous for pets and aside from
intoxication, can cause a coma or even death;
• Apple Seeds: can have varied effects;
• Apricot Pits: can cause respiratory difficulties such as breathing, coughing and sneezing;
• Avocado: can cause digestive problems. It is a very high salicylate that is poisonous to
cats and dogs. It will trigger fluid accumulation in the lungs and chest, leading to
difficulty breathing and death due to oxygen deprivation. Fluid accumulation can also
occur in the heart, pancreas and abdomen;
• Cherry Pits: can cause respiratory difficulties such as breathing, coughing and sneezing.
• Candy containing the sweetener Xylitol: can cause liver damage and even death;
• Mushrooms: contain toxins that will trigger numerous organ systems, including the
kidneys, liver and brain. Nervous system abnormalities, seizure, coma, vomiting, and
death can all result when a dog ingests mushrooms.
Generally, cooked and marinated foods should be avoided, as well as sauces and gravies,
which may contain ingredients that, although well tolerated by humans, may be toxic to
animals. Other miscellaneous foods that are toxic to dogs include: raw eggs and egg whites,
raw fish, nutmeg, salt, tobacco, trash items, persimmons, yeast and dough containing yeast,
liver, marijuana, hops, and human iron supplements. 461
6. EU rules on animal by-products
One of the first Directives of the European Community was the Council Directive
64/432/EEC. 462 This Directive applied to intra-Community trade in bovine animals and
458. Kobayashi K. (1981), "Onion poisoning in the cat", Feline Practice, 11 (1): 22-7; Stallbaumer M.
(1981), "Onion poisoning in a dog", Veterinary Record, 108 (24): 523-4; Harvey J. W., D. Rackear
(1985), "Experimental onion-induced hemolytic anemia in dogs", Veterinary Pathology, 22 (4): 387-92.
459. Eteng M. U., E. U. Eyong, E. O. Akpanyung, M. A. Agiang, C. Y. Aremu (1997), "Recent advances
in caffeine and theobromine toxicities: a review", Plant Foods for Human Nutrition, 51 (3): 231—243.
460. Tawde S. R, B. Puschner, T. Albin, S. Stump, R. H. Poppenga (2012), "Death by Caffeine:
Presumptive Malicious Poisoning of a Dog by Incorporation in Ground Meat", Journal of Medical
Toxicology, 8 (4): 436-440.
461. Kovalkovicova N., I. Sutiakova, J. Pistl, V. Sutiak (2009), ibid., 169-176.
462. Council Directive of 26 June 1964 "on animal health problems affecting intra-Community trade
in bovine animals and swine" (64/432/EEC) (Official Journal L 121, 29.7.1964, p. 164-184). This
125
Raffaella Leoci
swine for breeding, production or slaughter. Each Member State could ensure that only-
bovine and swine, which fulfill the general conditions, was sent from its territory to that of
another Member State. Bovine and swine covered by this Directive could show no clinical
sign of disease on the day of loading; had to have been obtained from a holding which was
situated in the center of an epizootic free area for at least three months prior to consignment;
and had been free from foot-and-mouth disease and bovine brucellosis, swine fever and con-
tagious porcine paralysis (Teschen disease) in the case of swine.
In 1982 the EC Commission, considering the appearance of certain contagious animal
diseases, noted a possible risk to the Community herd, as a result of intra-Community trade
and in the light of the experience gained from several 463 notifications. Adaptation to techni-
cal needs with a procedure calling for close cooperation between the Member States was put
into effect and adopted by the Directive 82/894/EEC. 464 Annex I of this Directive reported
the diseases which were the subject of the notification: foot-and-mouth disease, rinderpest
(cattle plaque), contagious bovine pleuropneumonia, bluetongue, swine vesicular disease,
classical swine fever, African swine fever, Teschen disease (contagious swine paralysis), fowl
plaque, and Newcastle disease. The ABPs were not yet considered.
In 1985 the first case of Bovine Spongiform Encephalopathy (BSE) was identified in
GB. In 1999 the Belgian dioxin 465 crisis was related to chicken feed. Also in 1999 there
was the kaolin scandal that involved high level of dioxin added to animal feed in Austria
and Germany. 466 Classical swine fever hit in 2000, 467 followed by foot and mouth disease
(FMD). 468 These crises, concerning human food and animal feed, exposed weaknesses in
Directive had been many times amended; the last amendments are Directives 97/12 and 98/46.
463. I.e. article 9 of Council Directive 64/432/EEC of 26 June 1964 on animal health problems af-
fecting intra-Community trade on bovine animals and swine, as last amended by Council of Directive
80/1274/EEC; Article 11 Council Directive 71/118/EEC of 15 February 1971 dealing with health
problems affecting trade in fresh poultry meat, as last amended by Council Directive 80/216/EEC;
Article 7 of Council Directive 72/461 /EEC of 12 December 1972 on health problems affecting intra-
Community trade in fresh meat, as last amended by Council Directive 8071099/EEC and Article 7
of Council Directive 80/215/EEC of 22 January 1980 on animal health problems affecting intra-
Community trade in meat products, as last amended by Council Directive 80/1 100/EEC.
464. Council Directive 82/894/EEC of 21 December 1982 on the notification of animal diseases
within the Community (Official Journal of the European Union, L378 of 31.12.82).
465. Dioxin was cited, in the European press for the first time, in relation to the accident at the
chemical plant of Hoffmann-LaRoche in Seveso, Italy. Due to an explosion a great amount of dioxin
was spread over the city of Seveso in 1976. In 1999, Belgium had a dioxin crisis caused by dioxin-
contaminated feed being fed to livestock. The source of the contamination was a Belgian at-rendering
company, where transformer oil with high levels of polychlorinated biphenyls (PCBs) and dioxins was
used to manufacture animal foods sale of Belgian poultry and eggs and all food items containing more
than 2% egg product.
466. Kaolinite is used, as 3% in animal feed, to improve the flow during pumping when moving the
animal feed from one store to another. German kaolin with high level of dioxin had also been added to
animal feed in Austria and Germany. In June 1999 animal feedings with added kaolinite were found
to have 1,5 to 30 pg i-TE/g resulting in a contamination of German turkeys of 30,6 pg i-TE/g fat.
467. Outbreaks have occurred in 1997-98 in the Netherlands, Germany, Belgium, Spain, Austria, and
Switzerland, costing an estimated $ 2.3 billion; 12.4 million animals were slaughtered. In 2000 in the
United Kingdom and in 2001 in Germany, Slovakia, Spain, and Romania.
468. Foot and Mouth Disease (FMD) or hoof-and-mouth disease (Aphthae epizooticae) is an infec-
126
Animal by-products (ABPs): origins, uses, and European regulations
the design and application of food legislation within the EU and lead to the Commission
including the promotion of a high level of food safety among its policy priorities. Over the
next years, the European Community adopted the Council Directives 90/425/EEC 469 and
90/667/EEC. 470
According to Council Directive 90/425/EEC, in the Member State of origin, the com-
petent authority must verify, among other things, that the animals and products for trade:
• meet the requirements of the relevant Directives detailed in Annex A (Council Di-
rectives 64/432/EEC, 88/407/EEC, 89/556/EEC, 90/426/EEC, 90/429/EEC) and
comply with the animal health requirements of the Member State of destination
(sheep and goats, live poultry, domestic rabbits, waste, hatching eggs);
• come from centers or organizations which are subject to regular official veterinary
checks;
• are accompanied by health certificates and other appropriate documents during transport;
• do not originate from holdings or regions which are subject to restrictions applying
to those animals or products because of the suspicion or existence of certain diseases;
• come from a country or region offering sufficient health guarantees from the point of
view of the country of destination;
• are transported in accordance with the hygiene rules in force.
Additionally, in the country of origin, a competent authority must carry out checks in
holdings, markets and assembly centers to verify that the live animals and products meet
European standards, especially as regards identification. Measures would be taken against
suppliers or consignors of animals and products who fail to comply with these rules.
Once the transport operation was completed he consignees of animals and products
dispatched from another Member State were responsible for those animals and products
upon their arrival at the destination and afterwards. If necessary, live animals had to be
quarantined at the place of destination or in a quarantine station. Checks had to be carried
out at the places where live animals and products from third countries could be brought into
EU territory, such as ports, airports and frontier posts with third countries. If there was an
outbreak of zoonosis, disease or any other risk to animal or human health, the Member State
of dispatch had to take the appropriate preventive and control measures, including restric-
tions on movement if the risk was serious.
tious and sometimes fatal viral disease that affects cloven-hoofed animals, including bovids. An out-
break of FMD, Type O, in Uruguay was reported on 27 October 2000. Alert measures in Uruguay had
been in place since the August 2000. In the same period FMD outbreaks in Argentina (type A24) and
Brazil (type O). An outbreak of FMD in Egypt was reported on 15 September 2000. In 2001, a seri-
ous outbreak of FMD in Britain resulted in the slaughter of many animals. Refer for more news: Tully
D. C, M. A. Fares (2008), "The tale of a modern animal plague: tracing the evolutionary history and
determining the time-scale for foot and mouth disease virus", Virology, 382 (2): 250—256.
469. Council Directive 90/425/EEC of 26 June 1990 concerning veterinary and zootechnical checks
applicable in intra-Community trade in certain live animals and products with a view to the comple-
tion of the internal market (Official Journal of the European Union, No L 224 of 18.8.90).
470. Council Directive 90/667/EEC of 27 November 1990 laying down the veterinary rules for the
disposal and processing of animal waste, for its placing on the market and for the prevention of patho-
gens in feedstuffs of animal or fish origin and amending Directive 90/425/EEC (Official Journal of the
European Union, No L 363 of 27.12.90).
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Raffaella Leoci
According to Article 3 of Council Directive 90/667/EEC, the following high-risk ma-
terial could be processed in a high-risk processing plant approved by the Member State or
disposed of by burning or burial:
• all bovine animals, pigs, goats, sheep, solipeds, poultry and all other animals kept for
agricultural production, which have died on the farm but were not slaughtered for
human consumption, including stillborn and unborn animals;
• dead animals not referred to in the previous point but which are designated by the
competent authority of the Member State;
• animals which are killed in the context of disease control measures either on the farm
or in any other place designated by the competent authority;
• animal waste including blood originating from animals which show, during the vet-
erinary inspection carried out at the time of slaughtering, clinical signs of diseases
communicable to man or other animals;
• all those parts of an animal slaughtered in the normal way which are not presented for
post mortem inspection, with the exception of hides, skins, hooves, feathers, wool,
horns, blood and similar products;
• all meat, poultry meat, fish, game and foodstuffs of animal origin which are spoiled
and thus present a risk to human and animal health;
• animals, fresh meat, poultry meat, fish, game and meat and milk products, imported
from third countries, which in the course of the inspections provided for in Commu-
nity legislation fail to comply with the veterinary requirements for their importation
into the Community, unless they are re-exported or their import is accepted under
restrictions laid down in Community provisions;
• without prejudice to instances of emergency slaughtering for reasons of welfare, farm
animals which have died in transit;
• animal waste containing residues of substances which may pose a danger to human
or animal health; milk, meat or products of animal origin rendered unfit for human
consumption by the presence of such residues;
• fish which show clinical signs of diseases communicable to man or to fish.
The competent authorities had where necessary to decide that high-risk material had to
be disposed of by burning or by burial where:
• transport to the nearest high-risk material processing plant of animals infected or sus-
pected of being infected with an epizootic disease was rejected because of the danger
of propagation of health risks;
• animals are infected with or suspected of being infected with a serious disease or
contain residues which could constitute a risk to human or animal health and which
could survive inadequate heat treatment;
• a wide-spread epizootic disease lead to a lack of capacity at the high-risk material
processing plant;
• animal waste concerned originated from places with difficult access;
• quantity and the distance to be covered did not justify collecting the waste.
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Animal by-products (ABPs): origins, uses, and European regulations
Burial had to be deep enough to prevent carnivorous animals from digging up the ca-
davers or waste and had to be in suitable ground so as to prevent contamination of water ta-
bles or any environmental nuisance. Before burial, the cadavers or waste had to be sprinkled
as necessary with a suitable disinfectant authorized by the competent authority.
Low-risk material had to be processed in a high-risk or low-risk processing plant ap-
proved in accordance with the present law, in a petfood plant, or in a plant preparing phar-
maceutical or technical products, or be disposed of by burning or burial in accordance with
the present law.
In addition to the animal waste referred to in Article 2 (low-risk material - animal
waste other than that covered by Article 3, which does not present serious risks of spreading
communicable diseases to animals or man), the following had to be deemed to be low-risk
material:
• products excluded under Article 3 (all those parts of an animal slaughtered in the
normal way which are not presented for post mortem inspection, with the exception
of hides, skins, hooves, feathers, wool, horns, blood and similar products), in so far as
they are used in the manufacture of feeding stuffs;
• fish caught in the open sea for the purposes of fishmeal production;
• fresh fish offal from plants manufacturing fish products for human consumption.
The mixture of low-risk material processed together with high-risk material had to be
deemed to be high-risk material.
Where low-risk material was processed in a petfood plant or a plant preparing pharma-
ceutical or technical products, the competent authority may require that it was dispatched,
stored and processed in a specific location and under specific conditions.
Fishmeal from industries that received and manufactured exclusively low-risk materials
intended for the manufacture of fishmeal had to meet the requirements laid down in Annex
II, Chapter III. 471
The products concerned should be subject to the rules for veterinary checks and any
protective measures laid down by Council Directive 90/425/EEC concerning veterinary
and zootechnical checks applicable in intra-Community trade of certain live animals and
products with a view to the completion of the internal market.
471. Requirements concerning the products after processing.
1. In the case of high-risk materials, samples of the finished products, taken directly after heat treat-
ment, must be free from heat-resistant pathogenic bacteria spores (Clostridium perfrigens absent in 1
g of the product).
2. Samples of the final products from both low-risk and high-risk material taken during or upon with-
drawal from storage at the processing plant must comply with the following standards:
Salmonella: absence in 25 g: n = 5, c = 0, m = 0, M = 0; Enterobacteriaceae: n = 5, c = 2, m = 10, M
= 3 x 10$ in 1 g, where:
n = number of units comprising the sample;
m = threshold value for the number of bacteria; the result is considered satisfactory if the number of
bacteria in all sample units does not exceed m;
M = maximum value for the number of bacteria; the result is considered unsatisfactory if the number
of bacteria in one or more sample units is M or more;
C = number of sample units the bacterial count of which may be between m and M, the sample still
being considered acceptable if the bacterial count of the other sample units is m or less.
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The European directives still do not deal about dangers posed by the use of ABPs, but
efforts are by now closer.
In January 2000, the adoption of the "White Paper on Food Safety" 472 followed to these
directives, in order to integrate the animal by-products sector into the "farm to table" ap-
proach for food safety.
As was stressed at the Helsinki European Council on December 1999, particular at-
tention would be focused on improving quality standards and reinforcing systems of checks
throughout the food chain, from farm to table.
The White Paper on food safety was an important element in this strategy. The Com-
mission proposed a number of measures that would enable food safety to be organized in a
more coordinated and integrated manner. These included:
• the establishment of an independent European Food Authority with responsibility for
independent scientific advice on all aspects relating to food safety, operation of rapid
alert systems, and communication of risks;
• an improved legislative framework covering all aspects of food products "from farm to
table";
• greater harmonization of national control systems;
• dialogue with consumers and other stakeholders.
The Commission set out the general principles on which European food safety policy
should be based:
• a comprehensive, integrated approach throughout the food chain;
• a clear definition of the roles of all stakeholders in the food chain (feed manufacturers,
farmers and food operators, the Member States, the Commission, consumers);
• traceability of feed and food and their ingredients;
• a coherent, effective and dynamic food policy;
• risk analysis (comprising risk assessment, management and communication);
• scientific advice to the highest standards of independence, excellence and transparency;
• application of the precautionary principle in risk management.
Since these measures were not considered sufficient, the Commission, based on data
provided by each Member State, issued a paper on 20 November 2001. 473 They provided a
description of the disposal situation on the disposal and the processing and uses of animal
by-products across the Community. The paper covered:
• Processing and disposal routes;
• Trade/ export of processed animal protein and rendered fat;
472. Commission of the European Communities, White paper on food safety, COM(1999)719 final,
Brussels, 12 January 2000. See for more news: Lauterburg D. (2001), Food Law: Policy & Ethics, Cav-
endish Publishing Ltd., London, pp. 38-49; Spriggs J.,G. Isaac (2001), Food Safety and International
Competitiveness: The Case of Beef, CABI Publishing, Oxon (UK) - New York (USA), pp. 145-166;
Smulders F. J. M., J. D. Collins, Edts. (2002), Food Safety Assurance and Veterinary Public Health: Food
Safety Assurance in pre-harvest phase, Wageningen Academic Publishers, Nederlands, pp. 17-19.
473. Commission service paper on the processing, disposal and uses of animal by-products in Member
States, MEMO/01/378, Brussels, 20 November 2001.
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Animal by-products (ABPs): origins, uses, and European regulations
• Collection, transportation and rendering costs;
• Storage capacity for rendered products and costs;
• Incineration, co-incineration and small on-farm incineration;
• Burial and/or landfill;
• Biogas;
• Composting and use of processed animal protein as fertilizer;
• Disposal capacity in Member States.
Finally, in 2002, a Regulation 474 on animal by-products was adopted by the European
Parliament and the Council that introduced more stringent conditions throughout the food
and feed chains requiring safe collection, transport, storage, handling, processing, uses and
disposal of all ABPs.
That Regulation sets up a completely new approach. In the past, raw material of a lower
health standard than the one used for human food were permitted for use in animal feeds.
The practice of recycling dead animals and material unfit for human consumption
into the feed chain was the main factor in the spreading of the BSE and other food scan-
dals (dioxin crises and foot and mouth disease) . The Article 22 of the Regulation (EC) No
1774/2002 prohibited this practice.
The Regulation classified ABPs into three categories based on their potential risk to animals,
the public or to the environment and set out how each Category must or may be disposed.
Category 1 (Article 4) materials (i.e. ABPs presented the highest risk. These were "ani-
mals suspected of being infected by a TSE in accordance with Regulation (EC) No 999/200 1
or in which the presence of a TSE has been officially confirmed"; or scrapie, "animals other
than farmed animals and wild animals, including in particular pet animals, zoo animals and
circus animals"; "experimental animals"; "wild animals, when suspected of being infected
with diseases communicable to humans or animals" in which there is the presence of resi-
dues of prohibited substance e.g. hormone used for growth promotion or environmental
contaminants e.g. dioxins, PCBs, "catering waste from means of transport operating inter-
nationally") that must be completely disposed of as waste by incineration or landfill after
appropriate heat treatment. Category 1 material shall not be imported or exported except in
accordance with this Regulation or with rules laid down under the procedure referred to in
Article 33 (Where reference is made to this paragraph - "Regulatory procedure", Articles 5
and 7 of Decision 1999/468/EC shall apply) .
Category 2 (Article 5) materials. These are other high-risk materials including animal
by-products of the following description, or any material containing such by-products: ma-
nure and digestive tract content; all animal materials collected when treating waste water
from slaughterhouses; products of animal origin containing residues of veterinary drugs;
474. Regulation (EC) No 1774/2002 of the European Parliament and of the Council of 3 October
2002 laying down health rules concerning animal by-products not intended for human consump-
tion (Official Journal of the European Union, No L 273 of 10.10.2002). Amended and implement-
ed by Commission Regulations (EC) No 79/2005, (EC) No 92/2005, (EC) No 93/2005, (EC) No
2067/2005, (EC) No 181/2006, (EC) No 208/2006, (EC) No 209/2006, (EC) No 1192/2006, (EC)
No 2007/2006, (EC) No 1877/2006, (EC) No 185/2007, (EC) No 829/2007, (EC) No 832/2007,
(EC) No 1256/2007, (EC) No 1432/2007, (EC) No 1576/2007, (EC) No 399/2008, (EC) No
437/2008, (EC) No 523/2008, (EC) No 777/2008, (EC) No 129/2009).
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Raffaella Leoci
ABPs presenting a risk of contamination with other animal diseases (e.g. animals which die
on farm or are killed in the context of disease control measures on farm) and may be recycled
for uses other than feeds after appropriate treatment (e.g. biogas, composting, oleo-chemical
products, etc). Category 2 material shall be collected, transported and identified and shall be
directly disposed of as waste by incineration in an approved incineration plant or processed
in a approved processing plant or transformed in a biogas plant or in a composting plant or
disposed of as waste by burial in a landfill, etc.
Category 3 (Article 6) material. These are the lowest-risk materials, that include animal
by-products of the following description, or any material containing such by-products: parts
of slaughtered animals which are fit for human consumption but are not intended for human
consumption for commercial reasons; parts of slaughtered animals which are rejected as unfit
for human consumption but are not affected by any signs of diseases communicable to hu-
mans or animals; hides and skins, hooves and horns, pig bristles and feathers originating from
animals that are slaughtered in a slaughterhouse after undergoing ante-mortem inspection and
were fit, as a result of such inspection, for slaughter; blood obtained from animals other than
ruminants that are slaughtered in a slaughterhouse and after undergoing ante mortem inspec-
tion were found fit as a result of such inspection; former foodstuffs of animal origin, or former
foodstuffs containing products of animal origin other than catering waste, which are no longer
intended for human consumption for commercial reasons; and shells, hatchery by-products
and cracked egg by-products originating from animals which did not show clinical signs of any
disease communicable through that product to humans or animals, etc.
The Regulation extended the current ruminant intra-species recycling (cannibalism)
ban to other species. Porcine animal by-products could not be fed to pigs and poultry ani-
mal by-products could not be fed to poultry. However, derogation was provided for in the
case of fish and fur animals subject to strict controls by the competent authority (Article 22).
While the uses of catering waste in feed for pigs and poultry was not the focus of the
Regulation, it was of major concern to nearly all Member States. Hence the Member States
had agreed on a total ban on such feeding practices in the revised Council Directive on
Swine Fever 475 .
Such uses of former foodstuffs and restaurant kitchen waste containing meat products
were at the origin of a number of major animal disease epidemics, which had led to enor-
mous losses to the farming and non-farming community.
The three EU institutions agreed on the ban on intra-species re-cycling (cannibalism).
Because catering waste fed to pigs could contain porcine material, catering waste feeding
would be inconsistent with the ban on cannibalism. It was also not possible to establish clear
traceability for catering waste. The adopted regulation was flexible, permitting a temporary
relaxation of the ban on the use of Category 3 catering waste in feed (Article 19). It requires
that only animal by-products derived from animals fit for human consumption (Category
3) could be used for animal feed. In other words, the same health standards required by EU
legislation for human food were required for animal feed.
In order to guarantee that ABPs derived from animals unfit for human consumption
cannot enter the human food or animal feed chain, the following requirements had been
introduced:
475. Council Directive 2001/89/EC of 23 October 2001 on Community measures for the control of
classical swine fever.
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Animal by-products (ABPs): origins, uses, and European regulations
• complete separation during collection, transport, storage, handling and processing of
animal waste not intended for animal feed or human food (Article 7 and 8);
• complete separation of plants dedicated to feed production from plants processing
other animal waste destined to destruction (Article 16, etc.);
• stricter rules for traceability of animal by-products, including the control of move-
ments of BSE specified risk material by a record keeping system and accompanying
documents or health certificates, and visual markers for animal proteins and fats in-
tended for destruction (Article 7, 8, Chapter III, IV, etc.).
The Regulation introduced a set of controls, which was as strict as the control estab-
lished for the food industry.
The Commission prepared a series of transitional and permanent implementing meas-
ures in order to prepare for and facilitate a swift application of the Regulation on 1 May
2003. 476 The temporary transitional measures covered:
• Feeding catering waste to animals (Austria, Germany); 477
• Feeding used cooking oil to animals (Ireland, UK); 478
• To avoid cross-contamination, the Regulation requires total separation between plants
handling Category 1, 2 and 3 materials. Transitional measures have been agreed for
the separation of oleo-chemical plants (Belgium, Germany, Italy, Netherlands, Spain,
Sweden, UK); the separation of processing plants dealing with heat treatment of ma-
terials (France, Finland); and the separation of intermediate plants dealing with col-
lection, handling, temporary storage and dispatching (Finland, Italy);
• Low-capacity incinerators/co-incinerators (Finland, UK); 479
• Manure processing standards (Belgium, France, Finland, Netherlands);
• Composting standards (all Member States)
• Biogas standards (all Member States)
• Processing standards for mammalian blood (Germany, Italy, Spain, UK)
• Collection, transportation and transformation of former foodstuffs (all Member
States)
The implementing measures covered:
• A permanent derogation for intra-species recycling of fur animals (Finland and Esto-
nia and Latvia); 480
476. Jakobsson C, E. B. Sommer, P. De Clercq, G. Bonazzi, J. Schroder (2002), The policy implemen-
tation of nutrient management legislation and effects in some European Countries, a presentation held on
18th April 2002 in Gent, Belgium at the final Workshop of the EU concerted action Nutrient Man-
agement Legislation in European Countries NUMALEC.
477. Germany and Austria had been granted a 4-year transitional period permitting the continued use
of swill feeding.
478. Transitional measures for used cooking oil in animal feed allowed industry in the UK and Ireland to
adapt their business practices. The transitional measures for used cooking oil lasted on 31 October 2004.
479. Commission Decision 2003/327/EC (Expired on 31/12/2004).
480. 2003/324/EC: Commission Decision of 12 May 2003 as regards a derogation from the intra-
species recycling ban for fur animals under Regulation (EC) No 1774/2002 of the European Parlia-
ment and of the Council.
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Raffaella Leoci
• A permanent derogation for the feeding of endangered/protected species of necropha-
gous (carrion) birds (France, Greece, Italy, Portugal, Spain); 481
• Rules on the burial and burning of animal by-products (all Member States); 482
• Rules for low capacity incinerators/co-incinerators (all Member States); 483
• The introduction of technical amendments to the Annexes (all Member States). Re-
vised models of import health certificates and repealing of old legislation, which ap-
ply on 1 May 2004 (Regulation (EC) No. 668/2004). The new health certificates are
adapted to an electronic format, allowing for the speedy transfer of trade documents.
• Health conditions have been established for new products such as collagen, egg prod-
ucts, tri-calcium phosphate and flavouring innards. 484
6.1 Regulation (EC) 1069/2009 and Regulation (EU) 142/2011
In 2009 a new Regulation (EC) No 1069/09 485 passed into European law with the
intention of clarifying some of the issues raised by Regulation (EC) No 1774/02 and its
derogation and to give a clear framework based on a risk assessment philosophy. Commis-
sion Regulation (EU) No 142/ 1 1 486 was developed to provide clarity and detailed provisions
for the implementation of Regulation (EC) No 1069/09 and implementing Council Direc-
tive 97/78/EC as regards certain samples. Testori Coggi 487 reported that the main reasons
for changing the regulation were simplification and clarification of rules on environmental
issues in conjunction with the parliament's new waste directive. The new ABP Regulation
had also to reduce administrative burden on establishments and made science-based modi-
fications to product categorization.
The new EU ABPs Regulation 1069/2009 and Commission Regulation (EU) 142/201 1
481. European Commission: Decisions 322/2003 and 830/2005.
482. Commission Regulation (EC) No 811/2003 of 12 May 2003 implementing Regulation (EC) No
1774/2002 of the European Parliament and of the Council as regards the intra-species recycling ban
for fish, the burial and burning of animal by-products and certain transitional measures.
483. Commission Regulation (EC) No 808/2003 of 12 May 2003 amending Regulation (EC) No
1774/2002 of the European Parliament.
484. Commission Regulation (EC) No 668/2004 of 10 March 2004 amending certain Annexes to
Regulation (EC) No 1774/2002 of the European Parliament and of the Council, as regards the impor-
tation from third countries of animal by-products.
485. Regulation (EC) No 1069/2009 of the European Parliament and of the Council of 21 October
2009 laying down health rules as regards animal by-products and derived products not intended for
human consumption and repealing Regulation (EC) No 1774/2002 (Animal by-products Regulation)
(Official Journal of theEuropean Union L 300 of 14.1 1.2009).
486. Commission Regulation (EU) No 142/2011 of 25 February 2011 implementing Regulation
(EC) No 1069/2009 of the European Parliament and of the Council laying down health rules as re-
gards animal by-products and derived products not intended for human consumption and implement-
ing Council Directive 97/78/EC as regards certain samples and items exempt from veterinary checks at
the border under that Directive (Official Journal of the European Union L 54 of 26.2.201 1), amended
by Commission Regulation (EU) No 749/2011 of 29 July 2011, Commission Regulation (EU) No
294/20 1 3 of 1 4 March 20 1 3.
487. Paola Testori Coggi, Deputy director general of DC SANCO, the European Commission's (EC's)
Directorate General for Health and Consumer Affairs.
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Animal by-products (ABPs): origins, uses, and European regulations
came into force on 4 March 201 1 and are referred to as the "EU ABPs Regulation" and the
"EU Commission Regulation" respectively.
For most premises, the Regulations require a "covered space to receive animal by-
products". 488
Under EU ABPs Regulation 1069/2009 animal by-products can fall into one of three
categories that reflect the level of risk to public and animal health.
According that Regulation "animal by-product" means entire bodies or parts of animals,
products of animal origin, and other products obtained from animals, which are not intend-
ed for human consumption (article 3). These include: fallen stock on farms, wild animals
when they are suspected of being diseased , slaughterhouse waste, skins, feathers, blood, and
meat, fish, milk and eggs when they are interned for human consumption (articles 8, 9, 10).
More specifically, in other word:
• Category 1 material 489 is defined in article 8 of EU ABPs Regulation 1069/2009. High-
est risk and consists principally of material that is considered a TSE risk (Regulation
(EC) No 999/2001), and those parts of an animal considered most likely to harbor a
disease such as BSE. Pet animals, zoo and circus animals, and experimental animals (ar-
ticle 2(d) of Directive 86/609/EEC) are also classified in this Category due to the level
of veterinary drugs and residues they may contain. Wild animals may also be classified
as Category 1 material when they are suspected of carrying a disease communicable to
humans or animals. Catering waste from means of international transport (i.e. which
has come from outside the EU), ABPs collected during the treatment of waste water,
and ABPs containing residues of substances and environmental contaminants are listed
in Group B(3) of Annex I to Directive 96/23/EC. However if such residues exceed the
permitted level laid down by Community legislation, they are also Category 1;
• Category 2 material 490 is defined in article 9 of the same EU ABPs Regulation. Cate-
gory 2 material is also high risk. It includes fallen stock and manure and digestive tract
488. Annex IX, Section 1, point 1(b) of SANCO/7066/2010 Rev. 4 (POOL/D 1/201 0/7066/7066R4-
EN.doc), Commission Regulation of implementing Regulation (EC) No 1069/2009 of the European
Parliament and of the Council laying down health rules as regards animal by-products and derived
products not intended for human consumption and implementing Council Directive 97/78/EC as
regards certain samples and items exempt from veterinary checks at the border under that Directive.
489. According to Article 13 of EU Commission Regulation N° 142/11, the competent authority may
authorise the use of Category 1 material consisting of entire bodies or parts of dead animals contain-
ing specified risk material for the feeding, in feeding stations, to endangered or protected species of
necrophagous birds and other species living in their natural habitat, for the promotion of biodiversity
and, outside feeding stations, if appropriate without prior collection of the dead animals, to wild ani-
mals, referred to point 1 (a) of Section 2 of Chapter II of Annex VI (species of necrophagous birds in
the Member States), subject to compliance with the conditions set out in Section 3 of that Chapter
(The competent authority must be satisfied, on the basis of an assessment of the specific situation of
the species concerned and their habitat, that the conservation status of the species will be improved and
must identify in the authorisation, holdings or herds within a geographically defined feeding zone).
490. According to Article 13 of EU Commission Regulation N° 142/1 1, operators may feed Category
2 and 3 material to the following animals, provided that such material comes from animals which were
not killed or did not die as a result of the presence or suspected presence of a disease communicable to
humans or animals: zoo animals, fur animals, dogs from recognised kennels or packs of hounds, dogs
and cats in shelters, maggots and worms for fishing bait.
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content. Category 2 is also the default status of any ABP not defined in the EU ABPs
Regulation. These materials include ABPs collected during the treatment of waste
water, ABPs from establishments or plants processing Category 2 material, products
of animal origin which have been declared unfit for human consumption due to the
presence of foreign bodies in those products, products of animal origin other than
Category 1 material that are imported or introduced from a third country and fail
to comply with Community veterinary legislation for their import or introduction
into the Community, fetuses, oocytes, embryos and semen which are not destined for
breeding purposes, dead-in-shell poultry.
• Category 3 materials are defined in article 10 of the EU ABPs Regulation. They are
considered low risk. Category 3 materials includeparts of animals that have been
passed fit for human consumption in a slaughterhouse but which are not intended
for consumption. Category 3 also includes products of animal origin, or foodstuffs
containing products of animal origin which are no longer intended for human con-
sumption for commercial reasons or due to manufacturing or packaging defects or
other defects that do not pose a risk to public or animal health. These may include
heads of poultry, hides and skins, including trimmings and splitting thereof, horns
and feet, including the phalanges and the carpus and metacarpus bones, tarsus and
metatarsus bones of animals other than ruminants requiring TSE testing, pig bris-
tles, feathers, products of animal origin, or foodstuffs containing products of animal
origin which are no longer intended for human consumption, pet food and feeding
stuffs of animal origin or feeding stuffs containing animal by-products or derived
products which are no longer intended for feeding for commercial reasons, blood,
placenta, wool, feathers, hair, horns, hoof cuts and raw milk originating from live
animals that did not show any signs of disease communicable through that product
to humans or animals, shells from shellfish with soft tissue or flesh, hatchery by-
products, eggs, egg by-products, including egg shells, day-old chicks killed for com-
mercial reasons, catering waste other than as referred to in article 8.
EU ABPs Regulation requires that mixtures of different categories of animal by-prod-
ucts must assume the categorization of the highest risk animal by-product in the mixture.
Therefore a mixture containing categories 1, 2 and 3 would be treated as Category 1 material
(articles 8 g and 9 g).
One the most significant change that the EU ABPs Regulation introduced is the con-
cept of end point in the manufacturing of ABPs, beyond which the processed products are
no longer subject to the requirements of the EU ABPs Regulation due to the eliminated
potential risks via heat or chemical substances (Article 5). Consequently, certain finished
products are excluded from the scope of Regulation 1069/2009 to reduce operational bur-
dens such as labeling or record-keeping. This simplification provided v for the management
of small quantities. 491
491. The point 52) of "Whereas": "Certain establishments or plants which handle only small quanti-
ties of animal by-products which do not pose a risk to public and animal health should be allowed to
dispose of such by-products by means other than disposal in accordance with this Regulation, under
official supervision". According to point 55): "For materials typically sent in small quantities for re-
search, educational, artistic or diagnostic use, special conditions should be laid down to facilitate the
movement of such materials within the Community. Bilateral arrangements facilitating the control of
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Animal by-products (ABPs): origins, uses, and European regulations
According to Article 3 of EU Commission Regulation No 142/201 1 the following de-
rived products may be placed on the market, and imported, without restrictions:
a. biodiesel which fulfills the requirements for the disposal and use of derived products
set out in point 2(b) of Section 3 of Chapter IV of Annex IV; 492
b. processed petfood which fulfills the specific requirements for processed petfood set
out in point 7(a) of Chapter II of Annex XIII; 493
c. dogchews which fulfill the specific requirements for dogchews set out in point 7(b)
of Chapter II of Annex XIII; 494
d. hides and skins of ungulates which fulfill the specific requirements for the end point
of those products set out in point C of Chapter V of Annex XIII; 495
e. wool and hair which fulfill the specific requirements for the end point for those prod-
ucts set out in point B of Chapter VII of Annex XIII; 496
f. feathers and down which fulfill the specific requirements for the end point for those
products set out in point C of Chapter VII of Annex XIII; 497
g. fur which fulfills the special requirements for the end point for that product set out
in Chapter VIII of Annex XIII; 498
h. fish oil for the production of medicinal products which fulfills the special require-
ments for the end point for that product set out in Chapter XIII of Annex XIII; 499
materials moved between the Member States sharing a common border should be permitted under
special circumstances".
492. Materials resulting from processing in accordance with the biodiesel production process may be
in the case of biodiesel and of residues from the distillation of biodiesel, used as a fuel without restric-
tions under this Regulation (end point) and used for the production of derived products for applica-
tion to land, in the case of potassium sulphate.
493. Processed petfood which has been manufactured and packaged in the Union in accordance with
point 3 and which has been tested in accordance with point 5 or which has been subject to veterinary
checks in accordance with Directive 97/78/EC at a border inspection post, may be placed on the
market without restrictions.
494. Dogchews which have been manufactured and packaged in the Union in accordance with point 4
and which has been tested in accordance with point 5 or which have been subject to veterinary checks
in accordance with Directive 97/78/EC at a border inspection post, may be placed on the market
without restrictions.
495. Hides and skins of ungulates which pursuant to the decision of an operator are destined for pur-
poses other than human consumption, and which comply with the requirements of Regulation (EC)
No 853/2004 for raw materials for gelatine or collagen intended for use in food may be placed on the
market without restrictions. Hides and skins having undergone the complete process of tanning, 'wet
blue', 'pickled pelts', limed hides (treated with lime and in brine at a pH of 12 to 13 for at least eight
hours) may be placed on the market without restrictions.
496. Factory-washed wool and hair, and wool and hair which has been treated by another method
which ensures that no unacceptable risks remain, may be placed on the market without restrictions.
497. Feathers, parts of feathers and down which have been factory-washed and treated with hot steam
at 100 °C for at least 30 minutes may be placed on the market without restrictions.
498. Furs which have been dried at an ambient temperature of 18 °C for two days at a humidity of
55% may be placed on the market without restrictions.
499. Fish oil derived from the materials referred to in point A.2 of Section 3 of Chapter II of Annex X,
which has been de-acidified with a NaOH solution at a temperature of 80 °C or more and which has
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i. gasoline and fuels that fulfill the specific requirements for products from the multi-
step catalytic process for the production of renewable fuels set out in point 2(c) of
Section 3 of Chapter IV of Annex IV. 500
The following uses of ABPs and derived products are prohibited: 501
• feeding of terrestrial animals of a given species other than fur animals with processed
animal protein derived from the bodies or parts of bodies of animals of the same species;
• feeding of farmed animals other than fur animals with catering waste or feed material
containing or derived from catering waste;
• feeding of farmed animals with herbage, either directly by grazing or by feeding with
cut herbage, from land to which organic fertilizers or soil improvers, other than ma-
nure, have been applied unless the cutting or grazing takes place after the expiration
(and after at least 2 1 days) of a waiting period which ensures adequate control of risks
to public and animal health;
• feeding of farmed fish with processed animal protein derived from the bodies or parts
of bodies of farmed fish of the same species (Article 11).
The available system for the disposal and use of animal by-products vary with the Category
and are listed in articles 12 (Category 1), 13 (Category 2) and 14 (Category 3) of the EU ABPs
Regulation. In general the higher the risk Category the fewer are the options for use.
The detailed rules on use and disposal are found in the EU Commission Regulation No
142/11.
Article 16 of the EU ABPs Regulation permits member States to avail themselves of
certain derogations for the use and disposal of animal by-products. By way of derogation
from articles 12, 13, and 14, animal by-products may be used for research and other specific
purposes in accordance with article 17 (research and other specific purposes) in the case
of animal by-products referred to in article 1 8 (special feeding purposes) that are used for
special feeding purposes, such as in the case of animal by-products referred to in article 19
(collection, transport and disposal); and are disposed of in accordance with that article ( a
competent authority may authorize the disposal by burial of dead pet animals and equidae
by burning or burial on site or by other means under official supervision which prevent the
transmission of risks to public); and are disposed of or used in accordance with alternative
methods which have been authorized in accordance with article 20 (authorization of alterna-
tive methods) in the case of Category 2 and Category 3 materials; and if authorized by the
competent authority and used for the preparation and application to land of bio-dynamic
subsequently been purified by distillation at a temperature of 200 °C or more, may be placed on the
market for the production of medicinal products without restrictions.
500. The multi-step catalytic process for the production of renewable fuels may be in the case of gaso-
line and the other fuels resulting from the process, used as a fuel without restrictions. The multi-step
catalytic process for the production of renewable fuels may be, in the case of used clay from bleaching
and sludge from the pre-treatment process, disposed of by incineration or co-incineration, transformed
into biogas, composted or used for the manufacture of derived products referred to in Article 36(a)(i)
of Regulation (EC) No 1069/2009.
501. The document, from the European Parliament's Committee on the Environment, Public
Health and Food Safety, stops short of altering the status quo on cattle feed, pointing out that be-
cause cows are vegetarian they should not be fed so-called "processed animal proteins" (PAP). The
same goes for sheep.
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Animal by-products (ABPs): origins, uses, and European regulations
preparations as referred to in article 12-lc (the use of biodynamic preparations is allowed) of
Regulation (EC) No 834/2007; 502 and in the case of Category 3 material; and if authorized
by the competent authority are used for feeding to pet animals, in the case of ABPs, except
for Category 1 material, which arise in the course of surgical intervention on live animals or
during birth of animals on farm and, if authorized by the competent authority, disposed of
on that farm.
Article 20 of the EU ABPs Regulation provides for the authorization of alternative
methods for use and disposal of ABPs. The procedure for authorization of an alternative
method of use or disposal of ABPs or derived products may be initiated either by the Com-
mission or, following an application, by a Member State or by an interested party that may
represent several interested parties.
The guidance in the Section 1 of the EU ABPs Regulation (Title II - Obligations of
operators) explains the registration, transportation, documentation and record keeping re-
quirements for premises handling ABPs. Additionally the guidance explains under what
circumstances bacteriological sampling is required and how this should be done.
The procedures for loading and unloading animal by-products at EU ABPs Regulation
approved premises must be agreed upon with the inspecting officer prior to issue of the ap-
proval or registration, and must be included in the plant's "standard operating procedure"
or HACCP plan. They must be designed to ensure that ABPs are unloaded/loaded with
minimal risk to animal or public health. In general, ABPs should be unloaded/loaded inside
the building. With the agreement of an inspecting officer and a permit, certain ABPs could
be unloaded in outside yards in tightly controlled circumstances, providing the ABPs are
moved into the building without delay. If unloading outside is permitted, the plant's con-
struction and operations must meet the agreement of the inspecting officer and must ensure
the following:
• the premises must have an impervious yard, constructed in a manner that allows it to
be cleaned and disinfected, and with a fall to a foul drain;
• the vehicle must park on the impervious yard and unloading must be carried out as
quickly as possible;
• the receiving container is leak proof and is securely covered immediately after loading;
• the receiving container is stored on an impervious surface constructed in a manner
that allows it to be cleaned and disinfected, and with a fall to a foul drain;
• immediate clean-up procedures must be in place in case of spillage.
Before commencing operations, operators are required to notify the competent author-
ity of establishments or plants under their control which are active at any stage of the genera-
tion, transportation, handling, processing, storage, placing on the market, distribution, use
or disposal of ABPs and derived products (Article 23 - EU ABPs Regulation) in order to be
either approved or registered before commencing operations. This requirement is subject to
some exceptions. Article 24 (EU ABPs Regulation) lists the type of establishments/activities
that require approval before they can operate:
502. Council Regulation (EC) No 834/2007 of 28 June 2007 on organic production and labelling
of organic products and repealing Regulation (EEC) No 2092/91 (Official Journal of the European
Union, L 189 of 20.7.2007).
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a. Incinerators (high and low capacity);
b. Compost (except home composting), biogas/anaerobic digestion;
c. Processing plants;
d. Biodiesel plants;
e. Petfood plants: hermetically sealed, chews, heated, raw, including petfood manufac-
tured in domestic kitchens;
£ Handling plants: all plants that carry out sorting, cutting, chilling, freezing, salting
or other preservation process, removal of hides and skins, removal, hygienization or
pasteurization, sieving, etc.;
g. Storage plants: all plants that store raw material and send it out again in the same
state;
h. Storage of derived products, plants that take in certain derived products (those
intended to be disposed of in landfill or by incineration or to be used as fuel for
combustion, used as feed (excluding plants approved under Regulation (EC) No
183/2005 503 ), use for organic fertilizers/soil improvers);
i. Boilers using tallow as fuel;
j. Organic fertilizer/soil improver manufacturers.
The following activities require registrations unless they are approved as above:
a. Manufacturers using wool, hair, pig bristles, feathers, apiculture by-products, bone,
bone products, horns, horn products, hooves, hoof products, milk, milk-based prod-
ucts, milk-derived products, colostrum, colostrum-based products;
b. Manufacturers of blood, blood products, hides and skins and products thereof, tan-
ning activities, commercial game trophies, oleochemicals;
c. Collection centers;
d. Users of ABPs to feed zoo/circus animals or other wild animals or special feeding uses;
e. Research and diagnostic samples;
f. Mixers of components to manufacture organic fertilize rs/soil improvers;
g. Manufacturers of cosmetic products, active implantable medical devices, medical de-
vices, in vitro diagnostic medical devices, veterinary medicinal products, medicinal
products, handlers of intermediate products;
h. Other operators as landfill sites, pet cemeteries, operators who place untreated wool
or hair (from farms) on the market;
i. Operators who receive hides and skins from their own animals back from abattoirs;
j. Operators who handle material as trade samples or for exhibition or artistic uses.
k. Haulers and transporters.
503. Regulation (EC) No 183/2005 of the European Parliament and of the Council of 12 January
2005 laying down requirements for feed hygiene (Official Journal of the European Union L 35 of
8.2.2005).
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Animal by-products (ABPs): origins, uses, and European regulations
Activities that do not require ABP registration:
• Plants that are approved or registered in accordance with Regulation (EC) No.
852/2004 or Regulation (EC) No. 853/2004;
• Plants listed above as requiring approval;
• Activities where ABPs are generated by farms and other places where animals are
kept, bred or taken care of.
According article 6 of EU Commission Regulation No 142/2011, the competent au-
thority has to ensure that incineration and co-incineration of animal by-products and de-
rived products will only take place in incineration plants and co-incineration plants which
have been granted a permit in accordance with Directive 2000/76/EC or for plants not re-
quired to have a permit under the same Directive, in incineration and co-incineration plants
which have been approved by the competent authority to carry out disposal by incineration,
or disposal or recovery of animal by-products or derived products, if they are waste, by co-
incineration. Operators of incineration plants and co-incineration plants shall comply with
the general requirements for incineration and co-incineration set out in Chapter I of Annex
III. 504 Operators of high-capacity and low-capacity incineration and co-incineration plants
shall comply with the requirements of Chapter II 505 and Chapter III of Annex III. 506
In derogation from article 12 (Disposal and use of Category 1 material) and Article
14(c) (disposed of in an authorized landfill, following processing) of EU ABPs Regulation
No 1069/2009, the competent authority may authorize the disposal of the following Cat-
egory 1 and 3 materials in an authorized landfill:
a. imported petfood or petfood produced from imported materials, from Category 1
material referred to in article 8(c) (animal by-products derived from animals which
have been submitted to illegal treatment) of EU ABPs Regulation No 1069/2009;
504. Among the main requirements we report the following. Operators of incineration and co-inciner-
ation plants shall ensure that the hygiene conditions (ABPs and derived products must be disposed of
as soon as possible after arrival, animals must not have access to the plants, ABPs and derived products
that are awaiting incineration or co-incineration or to ash resulting from their incineration, there must
be total physical separation between the incineration or co-incineration equipment and the livestock,
equipment must be dedicated entirely to the operation of the incinerator and not used elsewhere on
the holding, incompletely incinerated animal by-products must be reincinerated or disposed of by
other means) are met in the plants under their control.
505. Incineration or co-incineration plants treating only ABPs and derived products with a capacity of
more than 50 kg per hour (high-capacity plants) and which are not required to have a permit to oper-
ate in accordance with Directive 2000/76/EC shall comply with the following conditions: The plants
must be equipped for each line with at least one auxiliary burner that will be switched on automatically
when the temperature of the combustion gases after the last injection of combustion air falls below 850
°C or 1100 °C, as applicable. When ABPs or derived products are introduced into the incineration
chamber, the plant must operate an automatic system to prevent the introduction of ABPs until the
temperature of 850 °C or of 1 100 °C has been reached.
506. Incineration and co-incineration plants treating only ABPs and derived products with a maxi-
mum capacity of less than 50 kg of ABPs per hour or per batch (low-capacity plants) and which are not
required to have a permit to operate in accordance with Directive 2000/76/EC shall only be used for
the disposal of dead pet animals or Category 1 materials referred to in article 8 (b) , (e) and (f ) , Category
2 materials referred to in article 9 or Category 3 materials referred to in article 10 of Regulation (EC)
No 1069/2009.
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b. Category 3 material referred to in article 10(f) (products of animal origin, or food-
stuffs containing products of animal origin, which are no longer intended for human
consumption for commercial) and (g) (petfood and feedingstuffs of animal origin,
which are no longer intended for feeding for commercial reasons) of EU ABPs Regu-
lation No 1069/2009 (Article 7 of Commission Regulation (EU) No 142/201 1).
According of article 10 of EU Commission Regulation No 142/2011, operators shall
ensure that establishments and plants under their control comply with the following require-
ments for the transformation of ABPs and derived products into biogas or for composting:
• the requirements applicable to biogas and composting plants set out in Chapter I
(requirements applicable to plants);
• the hygiene requirements applicable to biogas and composting plants set out in
Chapter II (hygiene requirements applicable to biogas and composting plants);
• the standard transformation parameters set out in Section 1 of Chapter III (Standard
transformation parameters);
• the standards for digestion residues and compost set out in Section 3 of Chapter III
(standards for digestion residues and compost).
The competent authority will only approve biogas and composting plants if they comply
with the requirements laid down in Annex V of EU Commission Regulation No 1 42/20 1 1 .
A biogas plant must be equipped with a pasteurization/hygienisation unit, which cannot be
by-passed for the animal by-products or derived products introduced with a maximum par-
ticle size of 12 mm before entering the unit. The plant must also have equipment for moni-
toring that the temperature of 70 °C is reached within one hour and has recording devices
and a system to prevent insufficient heating. A composting plant must be equipped with a
closed composting reactor or closed area which cannot be by-passed for the ABPs or derived
products introduced into the plant, and it must be equipped with installations for moni-
toring temperature against time, recording devices to record the results of the monitoring
measurements of temperature, and an adequate safety system to prevent insufficient heating.
When a plant receives or dispatches animal by-products, ideally the vehicle will enter
the building for unloading or loading. This seems the only acceptable method for many by-
products such as slaughter and butchery waste, or where by-products are tipped onto the
floor or into a hopper.
According to article 20 of EU Commission Regulation (Requirements concerning certain
registered establishments and plants handling animal by-products and derived products):
1. Operators of registered plants or establishments or other registered operators shall
handle ABPs and derived products under the conditions set out in Chapter IV 507
507. f . Operators of registered plants or establishments or other registered operators shall handle ani-
mal by-products and derived products under the following conditions:
- premises must be constructed in a way permitting their effective cleaning and disinfection, where
appropriate;
- premises must have appropriate arrangements for protection against pests, such as insects, rodents
and birds;
- installations and equipment must be kept in hygienic condition, where necessary;
- animal by-products and derived products must be stored under conditions preventing contamination.
Operators shall keep records in a form which is accessible to the competent authority.
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Animal by-products (ABPs): origins, uses, and European regulations
of Annex IX (Requirements applicable to certain approved and registered establish-
ments and plants).
2. Registered operators transporting ABPs or derived products, other than between
premises of the same operator, shall in particular comply with the conditions set out
in point 2 of Chapter IV of Annex IX.
Paragraphs 1 and 2 shall not apply to:
a. approved operators who are transporting animal by-products or derived prod-
ucts as an ancillary activity;
b. operators who have been registered for transport activities in accordance with
Regulation (EC) No 183/2005.
Hides and skins are also treated as ABPs unless they are being used for the production of
gelatin and/or collagen for human consumption, 508 in which case they must have come from
animals that have passed ante and post mortem inspection and their storage must comply
with the requirements for fresh meat in the food hygiene legislation. They must also be kept
separate from hides and skins categorized as ABPs (Article 26 - EU ABPs Regulation). The
handling of hides and skins can be considered in a number of different circumstances:
• at abattoirs where animals under or over 48 months of age are slaughtered;
• at animal by-product plants that recover fallen stock hides where the fallen stock are
under or over 48 months of age;
• at stand-alone approved animal by-product hide stores or at collection centers and
tanneries.
Control of hides and skins derived from cattle over 48 months of age slaughtered for
human consumption and fallen cattle are subject to BSE testing. Hides of animals over 48
months of age can be recovered for use in the leather industry provided they have come from
animals that have been tested for Transmissible Spongiform Encephalopathy (TSE) with a
negative result. This includes hides from fallen stock over 48 months of age as well as those
slaughtered for human consumption.
Operators have to put in place, implement and maintain own checks in their estab-
lishments or plants in order to monitor compliance with this Regulation. They have to
ensure that no animal by-products or derived products suspected or discovered that do not
to comply with this Regulation leave the establishment or plant, unless destined for disposal
(Article 28 - EU ABPs Regulation).
Handling and storage plants require approval under the EU ABPs Regulation. They
must be adequately separated from other plants such as slaughterhouse and food handling
Registered operators transporting animal by-products or derived products, other than between prem-
ises of the same operator, shall in particular:
- have information at their disposal with regard to the identification of their vehicles, which allows
the verification of the use of the vehicles for the transport of animal by-products or derived products;
- clean and disinfect their vehicles, as appropriate;
- take all other necessary measures to prevent contamination and the spreading of diseases communi-
cable to humans or animals.
508. Paragraph 5, Chapter I, Section XV, Annex III of Regulation (EC) 853/2004 regarding the
requirements for collection centres and tanneries supplying hides and skins for the manufacture of
gelatine for human consumption (the same requirements apply for collagen).
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premises. They may receive ABPs from other premises, and all consignments of ABPs must
be accompanied by a commercial document.
ABPs and derived products destined for feeding to farmed animals, excluding fur animals,
may only be placed on the market provided they are or they are derived from Category 3 mate-
rial other than material referred to in article 10 (hides and skins, hooves, feathers, wool, horns,
hair and fur, (adipose tissue, and catering waste) (Article 31 - EU ABPs Regulation).
Organic fertilizers and soil improvers may be placed on the market and used provided
they are derived from Category 2 or Category 3 material. In addition, digestion residues
from transformation into biogas or compost may be placed on the market and used as
organic fertilisers or soil improvers (Article 32 - EU ABPs Regulation). Article 22 of EU
Commission Regulation states that the competent authority of the Member State where an
organic fertilizer or a soil improver, which has been produced from meat-and-bone meal
derived from Category 2 material or from processed animal protein, is to be applied to land
shall authorize one or more components which are to be mixed with those materials.
Cosmetic products, active implantable medical devices, in vitro diagnostic medical de-
vices, and veterinary medicinal products, may be placed on the market without restrictions
(Article 33 - EU ABPs Regulation). Pet food derived from Category 3 material may be
placed on the market (Article 35 - EU ABPs Regulation).
Operators may place on the market derived products, other than the products referred to
in articles 31 (ABPs and derived products destined for feeding to farmed animals, excluding
fur animals), 32 (Organic fertilizers and soil improvers, digestion residues from transfor-
mation into biogas or compost), 33 (cosmetic, active implantable medical devices, medical
devices, in vitro diagnostic medical devices, veterinary medicinal products) and 35 (petfood),
provided those products are not intended for use for the feeding to farmed animals or for
application to land from which such animals are to be fed or intended for feeding to fur
animals. Operators ensure the control of risks to public and animal health by safe sourcing
in accordance with article 37, safe treatment in accordance with article 38, and verifying that
the products are only used for safe end uses in accordance with article 39 where safe treatment
does not ensure sufficient control (Article 36 of EU ABPs Regulation). Article 24 of the EU
Commission Regulation confirms that the use of Category 1 material for the manufacture of
derived products that are intended to be ingested by or applied to humans or animals, other
than for derived products referred to in Articles 33 and 36 of EU ABPs Regulation, shall be
prohibited. Article 25 of the EU Commission Regulation states that the importation into and
the transit through the Union of the following ABPs shall be prohibited: unprocessed ma-
nure, untreated feathers and parts of feathers and down, beeswax in the form of honeycomb.
Wool and hair which has been factory-washed and furs which have been dried at an ambient
temperature of 18°C for a period of at least two days at a humidity of 55% can be imported
or may transit through the Union without any animal health conditions.
As a consequence, factory-washed wool and wool which has been treated by another
method which ensures that no unacceptable risks remain, may be placed on the market
without restrictions. Only processed wool could be placed on the market; unprocessed wool
could only be sent to intermediate plants for storage and/or technical plants for sorting and/
or washing. "Unprocessed wool" or "untreated wool" is defined in the new EU Commission
Regulation as: "wool that has not undergone factory washing, been obtained from tanning,
or been treated by some other method that ensures that no unacceptable risks remain".
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The export of ABPs and derived products destined for incineration or landfill is pro-
hibited. The export of ABPs and derived products to third countries that are not members
of the OECD for use in a biogas or composting plant is prohibited (Article 43 - EU ABPs
Regulation).
Each Member State will draw up a list of establishments, plants and operators that have
been approved or registered in accordance with this Regulation within its territory (Article
47 - EU ABPs Regulation).
7. EU rules on waste from ABPs
Already mentioned at the beginning: that the unused ABPs have to be classified as waste
and then destined for disposal. Most European waste policy and guidance is based on EU
legislation that gives strong direction on waste issues to member states. The main instru-
ments of establishing law and policy are the Directives that specify the objectives that the
EU seeks to achieve on particular issues of waste management. It is necessary for member
states to implement their requirements within prescribed timescales. Let us recall the main
waste directive.
7.1 Waste Directive
Waste Framework Directive 75/442/ EEC 509
This was the original framework directive on waste which was amended by 91/156/
EEC, 91/92/EEC, and the Commission Decision 96/350/EC. The directive relates to waste
disposal and the protection of the environment from harmful effects caused by the disposal
of waste. In particular it aims to encourage the recovery and use of waste in order to conserve
natural resources.
Wastes are defined (Article 1) as "any substance or object in the categories set out in
Annex I (Ql - Production or consumption residues not otherwise specified below; Q2
- Off-specification products; Q3 - Products whose date for appropriate use has expired;
Q4...etc...; Q16 - Any materials, substances or products which are not contained in the
above categories) which the holder discards or intends or is required to discard". The scope
of this directive excludes certain categories of waste (Article 2: gaseous effluents emitted
into the atmosphere; radioactive waste; waste resulting from prospecting, extraction, treat-
ment and storage of mineral resources and the working of quarries; animal carcasses and
the following agricultural waste: fecal matter and other natural, non dangerous substances
used in farming; waste water, with the exception of waste in liquid form; decommissioned
explosives).
The directive required member states to appoint competent authorities to draw up waste
management plans (Article 7) and to develop an integrated network of regional facilities
(Article 5). The directive also established the requirements for licenses, registration of carriers
(Article 9), and the polluter pays principle (Article 15).
Animal carcasses and ABPs are excluded from the aim of this directive.
509. Council Directive 75/442/EEC of 15/7/1975 on waste (Official Journal of the European Com-
munities L 1 94 of 25.7.1975).
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Waste Framework Directive 2008/98/EC 510
This Directive repealed Directive 2006/12/EC of the European Parliament and of the
Council of 5 April 2006 regarding waste (the codified version of Directive 75/442/EEC as
amended), hazardous waste Directive 91/689/EEC, and the Waste Oils Directive 75/439/
EEC. It provides for a general framework of waste management requirements and sets the
basic waste management definitions for the EU.
In its premise this Directive states that "The first objective of any waste policy should be
to minimize the negative effects of the generation and management of waste on human health
and the environment. Waste policy should also aim at reducing the use of resources and favor
the practical application of the waste hierarchy. In its Resolution of 24 February 1997 511 on
a Community strategy for waste management, the Council confirmed that waste prevention
should be the first priority of waste management, and that re-use and material recycling should
be preferred to energy recovery from waste, where and insofar as they are the best ecologi-
cal options. It is therefore necessary to revise Directive 2006/12/EC 512 in order to clarify key
concepts such as the definitions of waste, recovery and disposal, to strengthen the measures
that must be taken in regard to waste prevention, to introduce an approach that takes into ac-
count the whole life-cycle of products and materials and not only the waste phase, and to focus
on reducing the environmental impacts of waste generation and waste management, thereby
strengthening the economic value of waste. Furthermore, the recovery of waste and the use of
recovered materials should be encouraged in order to conserve natural resources".
According to Article 2, from the scope of this Directive, to the extent that they are cov-
ered by other Community legislation, are excluded:
a. ABPs including processed products covered by Regulation (EC) No 1774/2002, ex-
cept those which are destined for incineration, landfilling or use in a biogas or com-
posting plant;
b. carcasses of animals that have died other than by being slaughtered, including ani-
mals killed to eradicate epizootic diseases and that are disposed of in accordance with
Regulation (EC) No 1774/2002.
The definition of "waste" 513 is much simpler than that of the Directive 75/442/EEC.
Within the scope of this Directive fall the parts above mentioned of ABPs which are destined
for incineration, landfilling or use in a biogas or composting plant. According to Article 4,
the following waste hierarchy will apply as a priority order in waste prevention and manage-
ment legislation and policy:
a. prevention;
b. preparing for re-use;
c. recycling;
d. other recovery, e.g. energy recovery;
e. disposal.
510. European Parliament and Council Directive 2008/98/EC of 19 November 2008 on waste and repeal-
ing certain directives (Official Journal of the European Communities L 312/3 of 22 November 2008).
511. Official Journal of the European Communities C 76, 11.3.1997.
512. Directive 2006/12/EC had amended Directive 75/442/EEC.
513. Article 3: 'waste' means any substance or object which the holder discards or intends or is required
to discard.
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For ABPs the recycling, energy recovery, and disposal are applicable. The recovered parts
may be regarded as being by-products because the conditions of articles 5 and 6 are met.
According to article 7, the measures designed to amend non-essential elements of this
Directive relating to the updating of the list of waste (known as CER codes) established by
Decision 2000/532/EC 514 will be adopted in accordance with the regulatory procedure with
scrutiny referred to in Article 39 (2). 515 The list of waste will include hazardous waste and
will take into account the origin and composition of the waste and, where necessary, the
limit values of concentration of hazardous substances. The list of waste will be binding as
regards determination of the waste which is to be considered as hazardous waste.
In the list of wastes, ABPs can be classified with the following codes:
0.2 Wastes from agricultural, horticultural, hunting, fishing and aquacultural primary pro-
duction, food preparation and processing;
02 01 02 - Animal tissue waste (primary production wastes) ;
02 02 02 - Animal tissue waste (wastes from the preparation and processing of meat, fish
and other foods of animal origin);
02 02 03 - Material unsuitable for consumption or processing (wastes from the prepara-
tion and processing of meat, fish and other foods of animal origin).
As can be seen, ABPs are not considered dangerous. In reality, in the case of ABPs clas-
sified in Category 1 and 2 materials, the same CERs equipped with asterisk 516 can be used,
or the following:
02 01 99* (Animal tissue waste);
02 02 99* (Animal tissue waste).
As in Article 9, in order to strengthen the re-use and the prevention, recycling and
other modes of waste recovery (and so ABPs), Member States may take legislative or non-
legislative measures to ensure that any natural or legal person who professionally develops,
manufactures, processes, treats, sells or imports products (producer of the product) has ex-
tended producer responsibility.
As in Article 22 (Bio-waste), 517 Member States are encouraged will take measures, as
appropriate, and in accordance with Articles 4 and 13, and to encourage:
514. Commission Decision 2000/532/EC of 3 May 2000 replacing Decision 94/3/EC establishing a
list of wastes pursuant to Article 1(a) of Council Directive 75/442/EEC on waste and Council Deci-
sion 94/904/EC establishing a list of hazardous waste pursuant to Article 1(4) of Council Directive
91/689/EEC on hazardous waste (Official Journal of the European Communities L 226 of 6.9.2000).
515. Where reference is made to this paragraph, Article 5(1) to (4) and Article 7 of Decision 1999/468/
EC shall apply, having regard to the provisions of Article 8 thereof. (Article 5.1: The Commission shall
be assisted by a regulatory committee composed of the representatives of the Member States and
chaired by the representative of the Commission).
516. According to the Annex of the Decision 2000/532/EC, "Any waste marked with an asterisk (*)
is considered as a hazardous waste pursuant to Article 1(4), first indent, of Directive 91/689/EEC on
hazardous waste".
517. "Bio-waste" (Article 3) means biodegradable garden and park waste, food and kitchen waste
from households, restaurants, caterers and retail premises and comparable waste from food processing
plants. The ABPs may be included among the bio-wastes.
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a. the separate collection of bio-waste with a view to composting and digestion of bio-waste;
b. the treatment of bio- waste in a way that fulfills a high level of environmental protection;
c. the use of environmentally safe materials produced from bio-waste.
Member States will require any establishment or undertaking intending to carry out
waste treatment to obtain a permit from the competent authority (Article 23).
As in Article 26, where the following are not subject to permit requirements, Member
States will ensure that the competent authority keeps a register of:
a. establishments or undertakings which collect or transport waste on a professional
basis;
b. dealers or brokers;
c. establishments or undertakings which are subject to exemptions from the permit
requirements pursuant to Article 24 (exemptions from permit requirements).
Annex I lists the disposal operations: 1. Deposit into or on to land (e.g. landfill, etc.);
2. Land treatment (e.g. biodegradation of liquid or sludgy discards in soils, etc.); 3. Deep
injection (e.g. injection of pumpable discards into wells, salt domes, or naturally occur-
ring repositories, etc.); 4. Surface impoundment (e.g. placement of liquid or sludgy discards
into pits, ponds or lagoons, etc.); 5. Specially engineered landfill (e.g. placement into lined
discrete cells which are capped and isolated from one another and the environment, etc.);
6. Release into a water body except seas/oceans; 7. Release to seas/oceans including sea-bed
insertion; 8. Biological treatment not specified elsewhere in this Annex which results in final
compounds or mixtures which are discarded by means of any of the operations numbered
1 to 12; 9. Physico-chemical treatment not specified elsewhere in this Annex which results
in final compounds or mixtures which are discarded by means of any of the operations
numbered 1 to 12 (e.g. evaporation, drying, calcination, etc.); 10. Incineration on land; 11.
Incineration at sea (This operation is prohibited by EU legislation and international conven-
tions); 12. Permanent storage (e.g. emplacement of containers in a mine, etc.); 13. Blending
or mixing prior to submission to any of the operations numbered 1 to 12; 14. Repackaging
prior to submission to any of the operations numbered 1 to 13; 15. Storage pending any of
the operations numbered 1 to 14 (excluding temporary storage, pending collection, on the
site where the waste is produced).
7.2 Landfill Directive
The target of the Directive 99/31 /EC 518 was to provide for measures, procedures and
guidance to prevent or reduce, as far a possible, the negative environment effects of landfill-
ing waste. It also aimed to reduce methane emissions and to harmonized the controls on
landfill throughout the European Union by setting targets for a reduction in the volumes of
biodegradable municipal waste going to landfills.
It defined the different categories of waste (municipal waste, hazardous waste, non-
hazardous waste and inert waste) and applied that definition to all landfills designed as waste
disposal sites for the deposit of waste onto or into land. Landfills were divided into three
classes (Article 4):
518. Council Directive 1999/31/EC of 26 April 1999 on the landfill of waste (Official Journal of the
European Communities L 182 of 16/07/1999).
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Animal by-products (ABPs): origins, uses, and European regulations
• landfills for hazardous waste (any waste which was covered by Article 1 (4) of Council
Directive 91/689/EEC of 12 December 1991 on hazardous waste);
• landfills for non-hazardous waste (any substance or object which was covered by
Directive 75/442/EEC);
• landfills for inert waste (waste that did not undergo any significant physical, chemical or
biological transformations. Inert waste does not dissolve, burn or physically or chemi-
cally react, biodegrade or adversely affect other matter with which it comes into contact
in a way likely to give rise to environmental pollution or harm human health. The total
leachability and pollutant content of the waste and the ecotoxicity of the leachate was
insignificant and did not endangered the quality of surface water and/or groundwater).
The Directive (Article 3, p. 2 ) did not apply to:
• the spreading of sludges on the soil (including sewage sludge and sludge resulting
from dredging operation and similar matter for the purpose of fertilization or im-
provement);
• the use of inert waste in landfills for redevelopment or restoration work, or for con-
struction purposes;
• the deposit of unpolluted soil or of non-hazardous inert waste resulting from pros-
pecting and extraction, treatment and storage of mineral resources and from the
operation of quarries;
• the deposit of non-hazardous dredge sludges alongside small waterways from which
they have been dredged and of non-hazardous sludges in surface water, including the
bed and its subsoil.
A standard waste acceptance procedure (Article 6) is laid down so as to avoid any risks:
• waste must be treated before being landfilled;
• hazardous waste within the meaning of the Directive must be assigned to a hazardous
waste landfill;
• landfills for non-hazardous waste must be used for municipal waste and for non-
hazardous waste;
• landfill sites for inert waste must be used only for inert waste;
• criteria for the acceptance of waste at each landfill class must be adopted by the Com-
mission in accordance with the general principles of Annex II (Waste acceptance
criteria and procedures).
The following wastes (Article 5, p. 3) are not to be accepted in a landfill:
a. liquid waste;
b. waste which, in the conditions of landfill, is explosive, corrosive, oxidizing, highly
flammable or flammable as defined in Annex III to Directive 91/689/EEC;
c. hospital and other clinical waste which is infectious;
d. used tires, with certain exceptions;
e. any other type of waste which does not meet the acceptance criteria laid down in
Annex II.
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In terms of environmental protection the directive contains detailed provisions for the
permit (articles 8 and 9), cost (Article 10), acceptance procedures (Article 11), control and
monitoring procedures (Article 12), closure, and after-care procedures (Article 13). The re-
quirements will apply immediately to all new landfills, and existing sites will have to be
brought up to the higher standards or will have to close. All current licenses will need to be
reviewed and there will be a transitional period, extending up to 2009, for bringing existing
sites up to the required standard..
7.3 Packaging and Packaging Waste Directive
The EU first introduced measures on packaging waste management of, in 1985, 519 but
it was not able to bring about the effective harmonization of national policies. Only some
EU Member States introduced measures on packaging and packaging waste management
with a view to reducing environmental impact. As a consequence, diverging national legisla-
tion appeared in several Member States. For these and other reasons, Member States and
economic operators approached the Commission to introduce comprehensive legislation on
packaging.
In 1992, the Commission came forward with a proposal for a Council Directive on
Packaging and Packaging Waste. Following a prolonged discussion in the European Parlia-
ment and the Council of Ministers, Directive 94/62/EC 520 was adopted.
This Directive's purpose is to harmonize national measures in order to prevent or reduce
the impact of packaging waste on the environment. It contains provisions on the prevention,
recovery, and recycling of packaging waste.
In 2004, the Directive was reviewed to provide criteria clarifying the definition of the
term "packaging" and increase the targets for recovery and recycling of packaging waste. It
established percentage targets for the recovery of packaging waste and the essential require-
ments that all packaging must meet.
In 2005, the Directive was revised again to allow new Member States transitional peri-
ods for attaining the recovery and recycling targets.
7.4 End of Life Vehicles Directive
The Directive 2000/53/EC 521 aims to prevent waste from vehicles and sets out measures
for the reuse, recycling, and other forms of recovery of end-of life vehicles and their compo-
nents. Preventive measures intend to reduce disposal of waste and improve the environmen-
tal performance of the economic operators involved in the life cycle of vehicles.
519. Council Directive 85/339/EEC of 27 June 1985 on containers of liquids for human consumption
(Official Journal of the European Communities No L 176 of 6.7.85).
520. Directive 94/62/EC of European Parliament and Council of 20 December 1994 on packag-
ing and packaging waste (Official Journal of the European Communities No L 365 of 31.12.1994).
Amended by Directive 2004/12/EC, Directive 2005/20/EC, and Regulation (EC) No 219/2009.
521. Directive 2000/53/EC of the European Parliament and of the Council of 18 September 2000 on
end-of life vehicles - Commission Statements (Official Journal of the European Communities No L
269 of 21. 10.2000).
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7.5 Waste Incineration Directive
The Waste Incineration Directive 2000/76/EC 522 aims to minimize environmental and
human health impact on the of air, land and water emissions caused by the incineration or
co-incineration of hazardous and non-hazardous waste. Co-incineration plants are installa-
tions whose main purpose is to produce energy or goods and which use waste as a regular or
additional fuel source.
The Directive sets stringent operating conditions, technical requirements and emission
limit values. All incineration or co-incineration plants must be permitted and the permit
lists the categories and quantities of hazardous and no n- hazardous waste that may be treated.
Strict rules are imposed on the process to retain the waste at a sufficient temperature to guar-
antee complete waste combustion.
7.6 Other Directives of some importance are:
• Directive 2002/96/EC of 27 January 2003 regarding waste electrical and electronic
equipment (WEEE);
• Directive 2006/21 /EC of the European Parliament and of the Council of 15 March
2006 on the management of waste from extractive industries and amending Direc-
tive 2004/35/EC;
• Directive 2006/66/EC of the European Parliament and of the Council of 6 Septem-
ber 2006 on batteries and accumulators and waste batteries and accumulators and
repealing Directive 91/157/EEC.
8. Italian rules on ABPs and on waste from ABPs
In Italy, the EU rulings on waste, particularly the Directive n. 2008/98/CE, were ab-
sorbed by the Legislative Decree n. 152/2006. 523 The Article 185, point 2b, excludes, from
the scope of that decree, "the ABPs, including processed products (e.g. meat and bone meal),
provided for by the Council Directive n. 1774/2002 (now Council Directive n. 1069/2009,
in force from March 4, 201 1, above mentioned), except those destined for incineration, for
disposal in landfills or for use in a biogas or composting plant". The Article 184-bis excludes
from the discipline of the same decree the "ABPs" when the following conditions occur:
• the by-product (substance or object) is originated from a production process whose
primary purpose is not the production of the same;
• the by-product will certainly be used in the same or another production cycle;
• the by-product will be used directly without any further treatment;
• further use is lawful, with no impact on the environment and health.
The Article 184-ter lays down that "a waste ceases to be waste when it is subjected to
a recovery operation, including recycling and preparing for re-use and meets the specific
criteria to be adopted in accordance with the following conditions:
522. Directive 2000/76/EC of the European Parliament and of the Council of 4 December 2000 on
the incineration of waste (Official Journal of the European Communities No L 332 , 28/12/2000).
523. Legislative Decree 3 April, 2006, n. 152, part IV, "Rules on waste management and reclamation
of contaminated sites" (G. U. n. 88 del 14 April 2006, S. O. n. 96/L), then amended several times.
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• substance or object is commonly used for specific purposes;
• a market or a demand exists for such a substance or object;
• substance or object fulfills the technical requirements for specific purposes and meets
the existing legislation and standards that are applicable to products;
• the use of the substance or object will not have negative impacts on the environment
or on human health".
However, the Regulation (EC) n. 1069/2009 524 contains several provisions that overlap
or are in addition to those of the Legislative Decree n. 152/06. The Article 4 of Regulation
lays down that "Member States shall ensure that an adequate system is in place on their ter-
ritory ensuring that animal by-products are collected, identified and transported without
undue delay and treated, used or disposed of in accordance with this Regulation". According
to the articles 12, 13 and 14, Categories 1, 2 and 3 materials, will be directly disposed of as
waste by incineration or co-incineration without prior processing or following processing,
by pressure sterilization if the competent authority so requires, and permanent marking of
the resulting material.
Category 1 material will be directly disposed of as waste by incineration without prior
processing or following processing, by pressure sterilization if the competent authority so
requires, and permanent marking of the resulting material. In the case of Category 1 mate-
rial other than material referred to in article 8 525 will be disposed of by pressure sterilization
processing, permanent marking of the resulting material and burial in an authorized landfill.
Category 1 material referred to in article 8(f), 526 will be disposed of by burial in an author-
ized landfill or used as a fuel for combustion with or without prior processing.
Category 2 material will be disposed of in an authorized landfill, following processing
by pressure sterilization and permanent marking of the resulting material or will be used
for the manufacturing of organic fertilizers or soil improvers to be placed on the market in
accordance with article 32 (marketable organic fertilizers and soil improvers) following pro-
cessing by pressure sterilization, when applicable, and permanent marking of the resulting
material or will be composted or transformed into biogas.
The Category 3 material will be used for the production of raw petfood, to be placed
on the market in accordance with article 35 (other derived products placed on the pet food
market); will be composted or transformed into biogas or will be used as a fuel for combus-
524. This Regulation, having the force of law in Italy, was increased by Legislative Decree 1 October,
2012 , n. 186, "Disciplina sanzionatoria per la violazione delle disposizioni di cui al regolamento (CE)
n. 1069/2009 recante norme sanitarie relative ai sottoprodotti di origine animale e ai prodotti derivati
non destinati al consumo umano e che abroga il regolamento (CE) n. 1774/2002, e per la violazione
delle disposizioni del regolamento (UE) n. 142/2011 recante disposizioni di applicazione del regola-
mento (CE) n. 1069/2009 e della direttiva 97/78/CE per quanto riguarda taluni campioni e articoli
non sottoposti a controlli veterinari in frontiera" (GU n. 255 del 31 ottobre 2012).
525. Animals suspected of being infected by aTSE in accordance with Regulation (EC) No 999/2001
or in which the presence of a TSE has been officially confirmed and animals killed in the context of
TSE eradication measures.
526. Animals and parts of animals, other than those referred to in article 8 or article 10, that died
other than by being slaughtered or killed for human consumption, including animals killed for disease
control purposes; foetuses; oocytes, embryos and semen which are not destined for breeding purposes;
and dead-in-shell poultry.
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Animal by-products (ABPs): origins, uses, and European regulations
tion with or without prior processing, or will be applied to land without processing in the
case of raw milk, colostrum and products derived therefrom, which the competent authority
does not consider to present a risk of any disease communicable through those products to
humans or animals.
Any operator, establishment, or plant that generates, transports 527 , handles, processes,
stores, places on the market, distributes, uses, or disposes ABPs or derived products must
be registered before commencing operations using a commercial document 528 or, when re-
quired by the EU ABPs Regulation or by a measure adopted in accordance with paragraph
6 by a health certificate. Commercial documents and health certificates accompanying ABPs
or derived products, during transport have to include information on the origin, the desti-
nation, and the quantity of such products, and a description of the animal by-products or
derived products and their marking, when such marking is required.
According to Article 21, operators will ensure that ABPs and derived products are ac-
companied during transport by a commercial document or, when required by this Regula-
tion, or by a health certificate. With information on the origin, the destination and the
quantity of such products, and a description of the ABPs or derived products and their
marking, when such marking is required by this Regulation.
Operators will collect, transport and dispose of Category 3 catering waste in accordance
with national measures foreseen in Article 13 529 of Directive 2008/98/EC (on waste).
According to Article 22, operators consigning, transporting or receiving animal by-
products or derived products will keep a record of consignments and related commercial
documents and health certificates. Operators will have systems and procedures in place to
identify the other operators to which their ABPs or derived products have been supplied and
the operators from whom they have been supplied.
This information will be made available to the competent authorities on request..
According to Article 23, with a view to registration, operators will before commencing
operations, notify the competent authority of any establishments or plants under their con-
trol which are active at any stage of the generation, transport, handling, processing, storage,
placing on the market, distribution, use or disposal of ABPs and derived products; provide
the competent authority with information on the Category of ABPs or derived products un-
527. The use of bulk trailers which have a partition across the width of the camion, known as "split
trailers", has an economic advantage to the industry by enabling the collection of different categories
of by-product in a single vehicle movement. The Regulation 142/201 1, Annex VIII Chapter II, states
that all necessary measures must be taken to ensure that consignments of animal by-products and
derived products are identifiable, kept separate and identifiable during collection where the animal
by-products originate and during transportation.
528. The commercial document could be in accordance with the model set out under Commission
Regulation (EU) No 255/2013 of 20 March 2013 amending, for the purposes of adaptation to sci-
entific and technical progress, Annexes IC, VII and VIII to Regulation (EC) No 1013/2006 of the
European Parliament and of the Council on shipments of waste (Official Journal of the European
Communities L 79 of 21.3.2013) and shall accompany ABPs and derived products.
529. Member States shall take the necessary measures to ensure that waste management is carried out
without endangering human health, without harming the environment and, in particular:
- without risk to water, air, soil, plants or animals;
- without causing a nuisance through noise or odours; and
- without adversely affecting the countryside or places of special interest.
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der their control and the nature of the operations performed using ABPs or derived products
as starting material.
The Chapter V - Collection, transport, identification and traceability - of the EU Com-
mission Regulation 142/201 1 lays down, in the article 17 (requirements regarding commer-
cial documents and health certificates, identification, the collection and transport of ABPs
and traceability), that the:
1 . Operators will ensure that ABPs and derived products:
a. comply with the requirements for collection, transport and identification set out
in Chapters I (collection and transport) 530 and II (Identification) 531 of Annex VIII;
b. are accompanied during transport by commercial documents and health certifi-
cates in accordance with the requirements set out in Chapter III (commercial
documents and health certificates) 532 of Annex VIII (Amended by the Commis-
530. As from the starting point in the manufacturing chain referred to in Article 4 (1) of Regulation (EC)
No 1069/2009, ABPs and derived products must be collected and transported in sealed new packaging
or covered leak-proof containers or vehicles. Vehicles and reusable containers, and all reusable items of
equipment or appliances that come into contact with ABPs or derived products, must be maintained in a
clean condition. Reusable containers must be dedicated to the carriage of a particular animal by-product
or derived product to the extent necessary to avoid cross-contamination. Packaging material must be
disposed of, by incineration or by other means in accordance with Union legislation. Other provisions
deal with the temperature conditions of the transport of ABPs, derogation for collection and transport of
Category 3 material comprising of milk, milk-based products and milk-derived products, and of manure.
531. All necessary measures must be taken to ensure that: consignments of ABPs and derived products
are identifiable and kept separate and identifiable during collection; consignments of ABPs and derived
products are dispatched from one Member State to another Member State in packaging, containers
or vehicles which are prominently and, at least for the period of transport, indelibly colour-coded for
displaying information. The label attached to the packaging, container or vehicle must clearly indicate
the category of the animal by-products or of the derived products and bear the following words visibly
and legibly displayed on the packaging, a container or vehicle, as applicable:
- in the case of Category 3 material, "not for human consumption";
- in the case of Category 2 material and derived products from Category 2 material, "not for animal
consumption";
- in the case of Category 1 material and derived products from Category 1 material where they are
destined for disposal, "for disposal only";
- the manufacture of petfood, "for manufacture of pet food only";
- the manufacture of a derived product, "for manufacture of derived products only. Not for human or
animal consumption or for application to land";
- in the case of gelatine produced from Category 3 material, "gelatine suitable for animal consumption";
- in the case of collagen produced from Category 3 material, "collagen suitable for animal consumption";
- in the case of raw petfood, "as pet food only";
- etc., etc.
532. During transportation, a commercial document or, when required by this Regulation, a health
certificate must accompany ABPs and derived products. As already mentioned earlier, the commercial
document could be in accordance with the model set out under Commission Regulation (EU) No
255/2013 of 20 March 2013 amending, for the purposes of adaptation to scientific and technical
progress, Annexes IC, VII and VIII to Regulation (EC) No 1013/2006 of the European Parliament
and of the Council on shipments of waste - Official Journal of the European Communities L 79/19 of
21.3.2013. It must be produced at least in triplicate (one original and two copies). The original must
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Animal by-products (ABPs): origins, uses, and European regulations
sion Regulation (EU) No 255/2013 of 20 March 2013).
2. Operators consigning, transporting or receiving ABPs or derived products will keep
records of consignments and related commercial documents or health certificates in
accordance with the requirements set out in Chapter IV (Records) 533 of Annex VIII.
The Directive 2008/98/EC on waste, in turn, lays down that the establishments, the
producers of hazardous waste and the establishments and undertakings which collect or
transport hazardous waste on a professional basis, or act as dealers and brokers of hazard-
ous waste, will keep a chronological record of the quantity, nature and origin of the waste,
and, where relevant, the destination, frequency of collection, mode of transport and treat-
ment method foreseen in respect of the waste, and will make that information available, on
request, to the competent authorities. For hazardous waste, the records will be preserved
for at least three years except in the case of establishments and undertakings transporting
hazardous waste that must keep such records for at least 12 months. Documentary evidence
that the management operations have been carried out will be supplied at the request of the
competent authorities or of a previous holder (Article 35).
The Article 190 of the Italian Legislative Decree n. 152/06 lays down that the public
and the private producers of hazardous and non-hazardous waste which carry out disposal
or recovery of waste and/or produce waste from the same activity, the traders of waste, those
who collect or transport special waste 534 , if they do not adopt the "Waste Traceability Sys-
tem" (WTS)' 535 must keep a stock book (record of loading and unloading). 536 The Article
193 lays down that the public bodies and the enterprises, which collect and transport waste,
if they do not adopt the WTS, must use a "formulary of identification" (a kind of "com-
mercial document") in which they must specify the name and address of the producer, the
holder and the receiver of the waste, the origin, amount and type of waste, the date and the
way forward for the transport. The formulary will be completed in four copies. Two copies
are used by the carrier and accompany the waste.
The Article 212 of the same decree lays down that companies engaged in the collection
and transportation of waste are obliged to be entered on the "National Registry of Environ-
accompany the consignment to its final destination. The receiver must retain it. The producer must
retain one of the copies and the carrier the other. Member States may require that proof of the arrival of
the consignments is provided by the TRACES system or by a fourth copy of the commercial document
which is sent back by the receiver to the producer. The sender and carrier must keep copies for at least
two years. Health certificates must be issued and signed by the competent authority.
533. The records shall contain a description of the animal species for Category 3 material and derived
products therefrom, destined for use as feed material, the quantity of the material. In the case of re-
cords kept by any person consigning ABPs the records shall contain the date, the name and the address
of the transporter and of the receiver. In the case of records kept by any person transporting ABPs the
records must contain the date, the place of origin of the material, the name and the address of the
receiver. In the case of records kept by any person receiving ABPs the records shall contain the date, the
name and the address of the transporter and the place of origin of the material.
534. According to the Italian Legislative Decree n. 1 52/2006, Article 1 84, the "special waste" are those
produced from agricultural activities, from industrial, craft, trade and services, from activities such as
demolition, construction and excavations, from care activities.
535. In Italian "Sistema di tracciabilita dei rifiuti" (SISTRI), not yet operating in Italy.
536. Register similar to that adopted for recording the VAT.
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Raffaella Leoci
mental Managers" (Albo Nazionale Gestori Ambientali). 537 It is not possible to carry out the
collection and transportation of waste without that registration.
The various cases which arise in practice are also regulated in Italy by the "Guidelines 538
for the application of EU ABPs Regulation 1069/2009", but not everything is clear. What
it is not clear?
The Article 6 (communication of vehicles and reusable containers) of the Guidelines,
point 1, lays down that any undertaking which carries ABPs and derived products . . . must
notify the Veterinary Service of the Local Health Authority (LHA) 539 in the area where the
company is registered/ recognized (headquarters). In addition, the list of vehicles and/or re-
usable containers placed under its control (model and license plate) and its variations must
also notify the Veterinary Service of the Local Health Authority. The second point refers to
the vehicles and/or reusable containers for the transport of ABPs and derived products. The
previous point, however, states that WHAT can not be used for the transport of live animals,
food, feed and waste.
The point 1 of the Article 7 (Identification mode of vehicles and containers) lays down
that the Veterinary Service of the LHA, upon receiving the notification referred to in Article
6, records in a register any vehicle or vessel used for the transportation of ABPs or derived
products so as to provide an identification code.
According to the point 1 of the Article 8 (commercial document), the transport of ABPs
and derived products must be accompanied by the Commercial Document referred to in
Annex VIII, Chapter III of the EU Commission Regulation 142/201 1. In order to establish
local needs within their territory, egions may authorize for the Category 1, 2 and 3 materials:
the use of a simplified Commercial Document. The choice of that option must be commu-
nicated to the Ministry of Health.
In the case of point 2,when ABPs and derived products are intended to be disposed of
as waste, the Commercial Document must be replaced by the documentation required by
environmental legislation, as in Article 10.
The point 1 of Article 9 (traceability/ register) states that the persons consigning, trans-
porting or receiving ABPs and/or derived products must keep record of consignments of Ar-
ticle 22 of EU ABPs Regulation 1069/2009 and related Commercial Documents or health
certificates. In the subsequent points indicate the cases that do not require the maintenance
of the register.
The point 1 of the Article 10 (method of disposal as waste - in accordance with envi-
ronmental - of Category 1, 2 and 3 materials) lays down that disposal as waste of ABPs and
derived products of Category 1, 2 and 3 materials must be made in the manner prescribed
by environmental legislation as regards means of transport, waste formulary, and registration
of waste or WTS in the following cases:
537. According to the Decree of the Ministry of the Environment 28 April 1998, n. 406 (Official
Gazette of the Italian Republic no. 276 of 25 November 1998).
538. Agreement between the Italian Government, the Regions and the Autonomous Provinces of
Trento and Bolzano on the document containing Guidelines for the application of the Regulation
(EC) No 1069/2009 of the European Parliament and of the Council of 21 October 2009 laying down
health rules as regards animal by-products and derived products not intended for human consumption
and repealing Regulation (EC) No 1774/2002.
539. In Italy called ASL (Azienda Sanitara Locale).
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Animal by-products (ABPs): origins, uses, and European regulations
1.1. in incineration or co-incineration plants approved in accordance with environ-
mental legislation (Article 6, point 1, letter a) of the Commission Regulation
(EU) 142/2011);
1.2. in an authorized landfill in accordance with environmental legislations, follow-
ing processing by pressure sterilization and permanent marking of the resulting
material, if they are Category 1 materials, other than those referred to in Article 8,
letter a), points i) and ii) and Category 2 materials.
1.3., 1.4., 1.5., 1.6., 1.7, in authorized landfills, the other points according to Article 1 0
of EU ABPs Regulation 1069/2009.
The point 2 of the same article also provides: "the ABPs or derived products of any
Category, including the dead animal carcasses, during any subsequent phases of collecting
from the place of production (storing, processing in facilities approved in accordance with
EU ABPs Regulation 1069/2009) are still considered ABPs and therefore are subject to the
requirements of the Regulations, including transport".
According to the Guidelines (point 1, Article 1 - Registration plants) "all activities
of production, transport, handling, processing, storage, marketing, distribution, use and
disposal of ABPs and derived products are subject to registration procedure, if authoriza-
tion is not required, in accordance with Article 24 (Approval of establishments or plants) 540
of the EU ABPs Regulation 1069/2009 or, in the case of plants that generate by-products,
if they have not already been approved or registered in accordance with Regulation No
853/2004." 541 Such registration (point 3) does not exempt the operator from the notifica-
tion, if the ABPs or the derived products represent feed materials.
The operator (point 4) makes the notification of the opening, change in ownership
or type of activity, cessation, closing of any activity subject to registration, at the LHA or
the Single Desk for Productive Activities (according to the procedure established by each
Region) where the business is located or in which he is resident (in the case the activity is
devoid of establishment, such as the activity of transport for third parties or brokerage, etc.).
The registration is made after the notification (in a manner determined by individual
Regions) 542 . The operator must declare that the activity meets the minimum requirements
established the EU ABPs Regulation 1069/2009 and the Commission Regulation (EU)
142/2011. The notification will be accompanied by a Technical report and a Planim-
etry of premises. Registration for the transport activity (Article 23 del ABPs Regulation
540. Operators shall ensure that establishments or plants under their control are approved by the com-
petent authority, where such establishments or plants carry out one or more of the following activities:
processing of ABPs by pressure sterilization; disposal, as waste, by incineration of ABPs and derived
products; disposal or recovery of ABPs and derived products, if they are waste, by co-incineration; use
of ABPs and derived products as fuel for combustion; manufacturing of pet food; manufacturing of
organic fertilizers and soil improvers; transformation of ABPs and/or derived products into biogas or
compost; handling of ABPs after their collection, by way of operations such as sorting, cutting, chill-
ing, freezing, salting, removal of hides and skins or of specified risk material; storage of ABPs.
541. Regulation (EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004
laying down specific hygiene rules for on the hygiene of foodstuffs (Official Journal of the European
Communities L 139 of 30.4.2004).
542. Each registered plant or operator is placed on a national list of the Ministry of Health edited by
the respective Region or LHA.
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Raffaella Leoci
1069/2009) only applies to companies whose business is to transport of ABPs and derived
products.
According to Article 2 (approval of establishments), the establishments or plants, where
activities are carried out according to Article 24 of the EU ABPs Regulation 1069/2009, are
subject to approval. The procedure for approval must comply with the provisions of Article
44 (Procedure for approval) of the EU ABPs Regulation 1069/2009. Operators, who wish
with the use of implants, to engage in the activities, provided for in Article 44 of the EU
ABPs Regulation 1069/2009, must submit an application for authorization and accordance
with procedures established by the Region. The authorization document must specify:
activity carried out;
• type of product generated;
• category (Articles 8, 9 and 10 of EU ABPs Regulation 1069/2009);
• identification number.
The following types of plants are excluded from authorization and registration if they are
subject to other national disposition:
• incineration or co-incineration (Italian Decree Lgs, May 11, 2005 n. 133 and Direc-
tive 2000/76/CE;
• approved landfills (Directive 1 999/3 1/CE;
• establishments or plants that generate by-products whose activities have been ap-
proved or registered according to EU laws on food hygiene;
• biogas or compost plants outbuildings on to the farm when introducing manure;
• biogas or compost plants outbuildings on to the farm when introducing waste from
kitchens and catering;
• biogas or compost plants outbuildings on to the dairy farms when they introduce
ABPs from the treatment and processing of milk.
8.1 Conclusions
Summarization of the the situation follows. The management of ABPs is not subject to
Italian Legislative Decree No 152/2006, except in cases when ABPs are destined for incin-
eration, for disposal in landfills, or for use in a biogas or composting plant. Some questions
arise immediately: when do the ABPs become waste? At the beginning of transport to the ul-
timate fate site for disposal/recovery? Or upon arrival at the plant? Moreover, if there is a stop
at an intermediate site (also a treatment site ), how should ABPs be stored and managed?
ABPs are normally produced at the slaughter centers. Some parts may be intended to go
to a recovery center (mentioned above) others slated for disposal.
If all ABPs are destined to go to a selection and recovery center, and are in the presence of
by-productsthat are excluded from the discipline of Italian Legislative Decree No 152/2006,
the transport company would not have the obligation to enrollment in the National Reg-
istry of Environmental Managers. According to the EU ABPs Regulation 1069/2009, the
Commission Regulation (EU) 142/2011 and the Guidelines, the company must utilize a
commercial document during transport. The vehicles used must be marked with the identi-
fication code received from the LHA responsible for the area.
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Animal by-products (ABPs): origins, uses, and European regulations
On the other hand, if they are considered waste and therefore destined for incineration,
for disposal in landfills, or for use in a biogas or composting plant, the carrier is required
to register with the National Registry of Environmental Managers (category 4 or 5, with
the indication of the codes CER of the materials to be transported), use the "identification
formulary" , and (until the WTS will be adopted) keep the "loading and unloading registry".
Vehicles must be carriage authorized by the Regional Section of the Register cited. Danger-
ous ABPs (e.g. in the case of animals suspected of being infected by aTSE in accordance with
Regulation (EC) No 999/200 1 or in which the presence of a TSE or other type of dangerous
diseases has been officially confirmed) must be listed in Category 5 and also possess a health
certificate. According to the Guidelines (point 2 of Article 8), it is not necessary to have the
commercial document expected by the EU ABPs Regulation 1069/2009 and Commission
Regulation (EU) 142/201 1 and their successive amendments. This view, however, seems in-
consistent with the provisions of point 2 of Article 10 of the same Guidelines that, as already
mentioned, lays down that "the ABPs or derived products of any category, including the
carcasses of dead animals, in eventual successive stages to the collection from the production
site (storing, processing in facilities approved by EU ABPs Regulation 1069/2009), are still
considered ABPs and, therefore, subject to the obligations under the Regulation, transport
included." If we follow these rules the transport operator would not have the obligation of
registration in the National Registry of Environmental Managers, despite the point 1 of the
Article 10 of the Guidelines.
The centers of selection and recovery (they can also be located near or in slaughter
plants) as already mentioned must be in compliance with the provisions of the EU ABPs
Regulation 1069/2009, the Commission Regulation (EU) 142/2011 and the Italian Guide-
lines. In particular they will hold the registration issued by LHA as a result of notification
by the operator.
Operators carrying out both activities for which recognition is provided for in Article
24 of the EU ABPs Regulation 1069/2009 (both those provided by the Italian Legislative
Decree n. 152/08), must ensure a permanent and absolute separation of the selection and
recovery. As regards to transport, the same vehicle cannot carry both ABPs, as waste destined
for disposal and ABPs or derived products intended for recovery and recycling. In other
words, it does not seem possible to request registration and its identification code from the
LHA and at the same time to ask the Italian Regional Section of the Registry of Environ-
mental Managers, which has territorial jurisdiction, for the registration and the subsequent
authorization of the same vehicle.
The Italian Court of Cassation, III Section penal, has intervened on the subject with
several judgments (e.g. n. 2710 of 15 December 201 1 and n. 25364 of 27 June 2012) ruling
that the ABPs are not subject to the regulations regarding waste and are exclusively subject
to EC Regulation n. 1774/2002, only if they are actually classified as by-products, according
to the article 184 bis of Italian Legislative Decree n. 152/2006. Otherwise, if the producers
discard the ABPs to send their to the disposal, they remain subject to the rules laid down by
the latter decree.
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160
Animal by-products (ABPs): origins, uses, and European regulations
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4.10 Intestines
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4.11 Lungs
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4.13 Ovaries
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PART II - Treatment and legislation on ABPs and waste
5. Collection and treatment plants for ABP
Albihna A., B. Vinneras (2007), "Biosecurity and arable use of manure and biowaste — Treatment
alternatives", Livestock Science, vol. 112 (3): 232-239.
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Biogas Workshop, October 2-4, 2003, University of Southern Denmark Esbjerg/Denmark.
Buyuksonmez E, R. Rynk, T. F. Hess, E. Bechinski (1999), "Occurrence, Degradation, and Fate of
Pesticides During Composting Part I: Composting, Pesticides, and Pesticide Degradation", Com-
post Science and Utilization, vol. 7 (4), 66-82.
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Pesticides During Composting Part II: Occurrence and Fate of Pesticides in Compost and Com-
posting Systems", Compost Science and Utilization, vol. 8 (1), 61-81.
Buyuksonmez E, S. Sekeroglu (2005), "Presence of pharmaceuticals and personal care products (PP-
CPs) in biosolids and their degradation during composting", Journal of Residuals Science & Tech-
nology, vol. 2 (1), 31-40.
Cope R. B. (2005), "Allium species poisoning in dogs and cats", Veterinary Medicine, vol. 100 (8): 562-6.
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remediation", Biocycle, vol. 36 (10): 53-56.
Gault G., P. Berny, G. Lorgue (1995), "Plants which are toxic for pets", Recueilde medecine veterinaire,
vol. 171 (2-3): 171-176.
Gunaseelan V. N. (1997), "Anaerobic digestion of biomass for methane production: a review", Biomass
and Bioenergy, vol. 13:83-114.
Harvey J. W., D. Rackear (1985), "Experimental onion-induced hemolytic anemia in dogs", Veterinary
Pathology, vol. 22 (4): 387-92.
Kirchmayr R., R. Scherzer, D. L. Baggesen, R. Braun, A. Wellinger (2003), Animal by-Products and
Anaerobic Digestion - Requirements of the European Regulation (EC) No 177412002, IEA Bioen-
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Center of Excellence.
Kovalkovicova N., I. Sutiakova, J. Pistl, V. Sutiak (2009), "Some food toxic for pets", Interdisciplinary
Toxicology, vol. 2 (3): 169-76.
Kobayashi K. (1981), "Onion poisoning in the cat", Feline Practice, vol. 1 1 (1): 22-7; Stallbaumer M.
(1981), "Onion poisoning in a dog", Veterinary Record, vol. 108 (24): 523-4.
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production potentials of by-products from meat-processing industry", Journal of Hazardous Ma-
terials, vol. 164 (1): 247-255.
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187
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Southampton (UK), pp. 483-94.
Salgado B. S., L. N. Monteiro, N. S. Rocha (201 1), "Allium species poisoning in dogs and cats", Jour-
nal of Venomous Animals and Toxins including Tropical Diseases, vol. 17 (1): 4-11.
Tawde S. N., B. Puschner, T. Albin, S. Stump, R. H. Poppenga (2012), "Death by Caffeine: Presump-
tive Malicious Poisoning of a Dog by Incorporation in Ground Meat" , Journal of Medical Toxicol-
ogy, vol. 8 (4): 436-440.
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6. EU rules on animal by-products (ABPs)
Jakobsson C, E. B. Sommer, P. De Clercq, G. Bonazzi, J. Schroder (2002), The policy implementation
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18th April 2002 in Gent, Belgium at the final Workshop of the EU concerted action Nutrient
Management Legislation in European Countries NUMALEC.
Lauterburg D. (2001), Food Law: Policy & Ethics, Cavendish Publishing Ltd., London, pp. 38-49
Smulders E J. M., J. D. Collins, Edts. (2002), Food Safety Assurance and Veterinary Public Health: Food
Safety Assurance in pre-harvest phase, Wageningen Academic Publishers, Nederlands, pp. 17-19.
SpriggsJ.,G. Isaac (2001), Food Safety and International Competitiveness: The Case of Beef CABI Pub-
lishing, Oxon (UK) - New York (USA), pp. 145-166;
Tully D. C, M. A. Fares (2008), "The tale of a modern animal plague: tracing the evolutionary his-
tory and determining the time-scale for foot and mouth disease virus", Virology, vol. 382 (2):
250-256.
188
Animal by-products (ABPs): origins, uses, and European regulations
EU and Italian rules
(in chronological order)
Council Directive 64/432/EEC of 26 June 1 964 on animal health problems affecting intra-Commu-
nity trade in bovine animals and swine (Official Journal L 121 of 29.7.1964).
Council Directive 71/118/EEC of 15 February 1971 dealing with health problems affecting trade in
fresh poultry meat (Official Journal of the European Union L 055 of 08/03/1971).
Council Directive 72/461 /EEC of 12 December 1972 on health problems affecting intra-Community
trade in fresh meat (Official Journal of the European Union L 302 of 31.12.1972).
Council Directive 75/442/EEC of 15/7/1975 on waste (Official Journal of the European Communi-
ties L 194 of 25.7.1975).
Council Directive 80/1099/EEC of 11.11.1980 (Official Journal of the European Communities L
325 of 1.12.1980)
Council Directive 82/894/EEC of 21 December 1982 on the notification of animal diseases within the
Community (Official Journal of the European Communities L 378 of 31/12/1982).
Council Directive 82/894/EEC of 21 December 1982 on the notification of animal diseases within the
Community (Official Journal of the European Union L 378 of 31.12.82).
Council Directive 85/339/EEC of 27 June 1985 on containers of liquids for human consumption
(Official Journal of the European Communities L 176 of 6.7.85).
Council Directive 90/425/EEC of 26 June 1990 concerning veterinary and zootechnical checks appli-
cable in intra-Community trade in certain live animals and products with a view to the comple-
tion of the internal market (Official Journal of the European Union L 224 of 18.8.90).
Council Directive 90/667/EEC of 27 November 1990 laying down the veterinary rules for the disposal
and processing of animal waste, for its placing on the market and for the prevention of pathogens
in feedstuffs of animal or fish origin and amending Directive 90/425/EEC (Official Journal of the
European Union L 363 of 27.12.90).
Directive 94/62/EC of European Parliament and Council of 20 December 1994 on packaging and
packaging waste (Official Journal of the European Communities L 365 of 31.12.1994).
Council Directive 97/78/EC of 18 December 1997 laying down the principles governing the organi-
sation of veterinary checks on products entering the Community from third countries (Official
Journal of the European Communities L 24 of 30.1.1998).
Decree of the Italian Ministry of the Environment 28 April 1998, n. 406 (Official Gazette of the Ital-
ian Republic no. 276 of 25 November 1998).
Council Directive 1999/31 /EC of 26 April 1999 on the landfill of waste (Official Journal of the Euro-
pean Communities L 182 of 16.7.99).
Commission Decision 2000/532/EC of 3 May 2000 replacing Decision 94/3/EC establishing a list
of wastes pursuant to Article 1 (a) of Council Directive 75/442/EEC on waste and Council Deci-
sion 94/904/EC establishing a list of hazardous waste pursuant to Article 1 (4) of Council Direc-
tive 91/689/EEC on hazardous waste (Official Journal of the European Communities L 226 of
6.9.2000).
Directive 2000/53/EC of the European Parliament and of the Council of 18 September 2000 on end-
of life vehicles - Commission Statements (Official Journal of the European Communities L 269
of21. 10.2000).
Directive 2000/76/EC of the European Parliament and of the Council of 4 December 2000 on the
incineration of waste (Official Journal of the European Communities L 332/9 1 of 28. 1 2.2000) .
189
Raffaella Leoci
Council Directive 2001/89/EC of 23 October 2001 on Community measures for the control of classi-
cal swine fever (Official Journal of the European Communities L 316 of 1.12.2001).
Commission service paper on the processing, disposal and uses of animal by-products in Member
States, MEMO/01/378, Brussels, 20 November 2001.
Regulation (EC) 1774/2002 of the European Parliament and of the Council of 3 October 2002 laying
down health rules concerning animal by-products not intended for human consumption (Official
Journal of the European Union L 273 of 10.10.2002).
Commission Decision 2003/324/EC of 12 May 2003 as regards a derogation from the intra-species
recycling ban for fur animals under Regulation (EC) No 1774/2002 of the European Parliament
and of the Council (Official Journal of the European Union L 117 of 13.5.2003).
Commission Regulation (EC) No 808/2003 of 1 2 May 2003 amending Regulation (EC) No 1 774/2002
of the European Parliament (Official Journal of the European Union L 1 17 of 13.5.2003).
Commission Regulation (EC) No 811/2003 of 12 May 2003 implementing Regulation (EC) No
1774/2002 of the European Parliament and of the Council as regards the intra-species recycling
ban for fish, the burial and burning of animal by-products and certain transitional measures (Of-
ficial Journal of the European Union L 1 17 of 13.5.2003).
Regulation (EC) No 2003/2003 of the European Parliament and of the Council of 13 October 2003,
relating to fertilisers (Official Journal of the European Union L 304 on 21.1 1.2003).
Directive 2004/12/EC of the European Parliament and of the Council of 1 1 February 2004 amending
Directive 94/62/EC on packaging and packaging waste - Statement by the Council, the Commis-
sion and the European Parliament (Official Journal of the European Union L 047 of 1 8.02.2004) .
Commission Regulation (EC) No 668/2004 of 10 March 2004 amending certain Annexes to Regula-
tion (EC) No 1774/2002 of the European Parliament and of the Council, as regards the importa-
tion from third countries of animal by-products (Official Journal of the European Union L 112
of 19.4.2004).
Regulation (EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004 lay-
ing down specific hygiene rules for on the hygiene of foodstuffs (Official Journal of the European
Communities L 139 of 30.4.2004).
Regulation (EC) No 183/2005 of the European Parliament and of the Council of 12 January 2005
laying down requirements for feed hygiene (Official Journal of the European Union L 35 of
8.2.2005).
Directive 2005/20/EC of the European Parliament and of the Council of 9 March 2005 amending
Directive 94/62/EC on packaging and packaging waste (Official Journal of the European Union
L 70 of 16.3.2005).
Legislative Decree 3 April, 2006, n. 152, part IV, "Rules on waste management and reclamation of
contaminated sites" (Official Gazette of the Italian Republic No 88 del 14 April 2006, S. O. n.
96/L).
Council Regulation (EC) No 834/2007 of 28 June 2007 on organic production and labelling of or-
ganic products and repealing Regulation (EEC) No 2092/91 (Official Journal of the European
Union L 189 of 20.7.2007).
European Parliament and Council Directive 2008/98/EC of 19 November 2008 on waste and repeal-
ing certain directives (Official Journal of the European Communities L 312/3 of 22 November
2008).
Regulation (EC) No 219/2009 of the European Parliament and of the Council of 11 March 2009
adapting a number of instruments subject to the procedure referred to in Article 25 1 of the Treaty
to Council Decision 1999/468/EC with regard to the regulatory procedure with scrutiny — Ad-
190
Animal by-products (ABPs): origins, uses, and European regulations
aptation to the regulatory procedure with scrutiny — Part Two (Official Journal of the European
Communities L 87 of 31.3.2009).
Regulation (EC) No 1069/2009 of the European Parliament and of the Council of 21 October 2009
laying down health rules as regards animal by-products and derived products not intended for
human consumption and repealing Regulation (EC) No 1774/2002 (Animal by-products Regu-
lation) (Official Journal of the European Union L 300 of 14.1 1.2009).
Commission Directive 2010/67/EU of 20 October 2010, amending Directive 2008/84/EC laying
down specific purity criteria on food additives other than colours and sweeteners (Official Journal
of the European Union L. 277 of 21.10.2010).
Directive 2010/75/EU of the European Parliament and of the Council of 24 November 2010 on in-
dustrial emissions (integrated pollution prevention and control) (Official Journal of the European
Union L 334 of 17.12.2010).
Commission Regulation (EU) No 142/2011 of 2 5 February 2011, implementing Regulation (EC) No
1069/2009 of the European Parliament and of the Council laying down health rules as regards an-
imal by-products and derived products not intended for human consumption and implementing
Council Directive 97/78/EC as regards certain samples and items exempt from veterinary checks
at the border under that Directive (Official Journal of the European Union L 54 of 26.02.201 1).
Commission Regulation (EU) No 749/201 1 of 29 July 201 1 amending Regulation (EU) No 142/201 1
implementing Regulation (EC) No 1069/2009 of the European Parliament and of the Council
laying down health rules as regards animal by-products and derived products not intended for
human consumption and implementing Council Directive 97/78/EC as regards certain samples
and items exempt from veterinary checks at the border under that Directive (Official Journal of
the European Union L 198 of 30.7.2011).
Legislative Decree 1 October, 2012 No 186, "Disciplina sanzionatoria per la violazione delle dispo-
sizioni di cui al regolamento (CE) n. 1069/2009 recante norme sanitarie relative ai sottopro-
dotti di origine animale e ai prodotti derivati non destinati al consumo umano e che abroga il
regolamento (CE) n. 1774/2002, e per la violazione delle disposizioni del regolamento (UE) n.
142/2011 recante disposizioni di applicazione del regolamento (CE) n. 1069/2009 e della diret-
tiva 97/78/CE per quanto riguarda taluni campioni e articoli non sottoposti a controlli veterinari
in frontiera" (Official Gazette of the Italian Republic No 255 del 31 ottobre 2012).
Commission Regulation (EU) No 294/2013 of 14 March 2013 amending and correcting Regulation
(EU) No 142/201 1 implementing Regulation (EC) No 1069/2009 of the European Parliament
and of the Council laying down health rules as regards animal by-products and derived products
not intended for human consumption and implementing Council Directive 97/78/EC as regards
certain samples and items exempt from veterinary checks at the border under that Directive (Of-
ficial Journal of the European Union L 98 of 6.4.2013).
Commission Regulation (EU) No 255/2013 of 20 March 2013 amending, for the purposes of ad-
aptation to scientific and technical progress, Annexes IC, VII and VIII to Regulation (EC) No
1 0 1 3/2006 of the European Parliament and of the Council on shipments of waste (Official Jour-
nal of the European Communities L 79 of 21.3.2013).
191
Raffaella Leoci
192
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